Engineered platelets for targeted delivery of a therapeutic agent

ABSTRACT

The present invention provides engineered platelets with chimeric platelet receptors (CPR) with a desired target specificity. Additionally, the engineered platelets may comprise cargo which may be released upon activation of the platelet. Additionally, the platelets may be generated in vitro from megakaryocytes engineered to generate non-thrombogemc platelets.

SEQUENCE LISTING

The present application is being filed along with a Sequence Listing inelectronic format. The Sequence Listing file, entitled SEQLIST.txt, wascreated on Dec. 14, 2020, and is 46,300 bytes in size. The informationin electronic format of the Sequence Listing is incorporated herein byreference in its entirety.

FIELD OF THE INVENTION

The invention relates to engineered platelets for targeted therapeuticdelivery systems.

BACKGROUND OF THE INVENTION

Platelets are small and enucleated and cannot divide or reproduce. Inthe human body, they perform the important function of recognisinginjured tissue and releasing their contents to reduce or preventbleeding. Thrombopoietin from the kidneys and liver contact a myeloidstem cell causing differentiation into a megakaryoblasorphant, andadditional signals result in differentiation of the megakaryoblast intoa progenitor megakaryocyte. Progenitor megakaryocytes are large cellswith platelet precursor extensions that bud off fragments as they divideand proliferate to create platelets.

Mitochondria, microtubules, and vesicles are contained within theplatelets, and the platelets have a life span of about 10 days beforeclearance by macrophages. Platelets have a volume of about 7 μm³ and adiameter of 300 nm. They are metabolically active and can alter geneexpression through post-transcriptional control of preloaded mRNAexpression (e.g. by miRNAs). On activation, granulation is stimulated toalter the shape and release the contents of the intracellular vesicles.

Platelets respond to a variety of extra cellular signals through adiverse set of signaling pathway receptors. Receptors act both totrigger intracellular signaling cascades resulting in plateletdegranulation and effector release and to cause platelet aggregation andadhesion. glycoprotein VI platelet (GPVI) signaling functionsanalogously to many immune cell receptors—such as the TCR.Interestingly, platelets also express toll-like receptors (TLRs) and canmediated targeted killing of bacteria via peptide secretion and immunesystem activation.

A huge variety of products are released on platelet degranulation.Vesicles are released by exocytosis. Platelets contain three primarysubtypes of vesicles: α-granules (50 to 80 per platelet), dense granules(3 to 8 per platelet), and large dense core vesicles (LDCV) (about10,000 per platelet). Different mutations can selectively disrupt thebiogenesis of each vesicle subtype.

α-granules have a diameter of about 200 to 500 nm and make up about 10%of the platelet's volume. Most effector proteins are found inα-granules. For example, effector proteins released from α-granulesinclude: integral membrane proteins, such as P-selectin, αIIbβ, andGPIbα; coagulants/anticoagulants and fibrinolytic proteins, such asfactor V, factor IX, and plasminogen; adhesion proteins, such asfibrinogen and von Willebrand Factor (vWF); chemokines, such as CXCL4(cytokine (C-X-C motif) ligand 4), also known as platelet factor 4 orPF4, and CXCL12 (cytokine (C-X-C motif) ligand 12), also known asstromal cell-derived factor 1 alpha or SDF-1α; growth factors, such aselongation growth factor (EGF) and insulin-like growth factor 1 (IGF);angiogenic factors/inhibitors, such as vascular endothelial growthfactor (VEGF), platelet-derived growth factor (PDGF), and angiostatins;and immune mediators, such as immunoglobulin G (IgG) and complementprecursors.

Dense granules have a diameter of about 150 nm and make up about 1% ofthe platelet's volume. Effector proteins released from dense granulesinclude cations, such as Ca²⁺ and Mg²⁺; polyphosphates; bioactiveamines, such as serotonin and histamine; and nucleotides, such asadenosine diphosphate (ADP) and adenosine triphosphate (ATP).

LDCVs have a diameter in the range of about 150 nm to about 300 nm andmake up about 13.5% of the platelet's volume. Effector proteins releasedfrom LDCVs include structural proteins (e.g., granins andglycoproteins); vascoregulators (e.g., cateholamines, vasostatins,renin-angiotensin); paracrine signaling factors (e.g., guanylin,neurotensin, chromogranin B); immune mediators (e.g., enkelytin andubiquitin); opiods (e.g., enkephalins and endorphins); ions (e.g., Ca²⁺,Na+, Cl—), and nucleotides and polyphosphates (e.g., adenosinemonophosphate (AMP), guanosine diphosphate (GDP),uridine-5′-triphosphate (UTP)).

Current cell therapies based on engineered chimeric antigen receptor Tcells (CAR-T cells) have shown promise treating cancer; however,concerns regarding their safety, specifically oncogenic transformationin the patient, and the limited ability to generate a generic oruniversal therapeutic product have restricted their use to a smallnumber of patients. There is a long felt need in the art for a new typeof therapy with the potential to treat cancer, autoimmune conditions,and infections, free from the safety, cost, and patient matching issueswhich plague current cell therapeutic products.

SUMMARY OF THE INVENTION

Various embodiments of the invention described herein provide a chimericplatelet receptor (CPR) comprising:

a) an intracellular domain that is a platelet stimulation domain andcomprises domains from an immunoreceptor tyrosine-based activation motif(ITAM) receptor; and

b) a heterologous targeting domain that recognizes and binds a target.

By a heterologous targeting domain we mean that the targeting domain isheterologous to the intracellular platelet stimulation domain i.e. thetargeting domain is not the usual extracellular domain associated withthe intracellular domain. The heterologous targeting domain may bind toan endogenous target, for example may bind to a tumour antigen that isendogenous to a subject but, by virtue of the CPR being chimeric, thetargeting domain is heterologous to the internal platelet stimulationdomain.

The invention described herein also provides a chimeric plateletreceptor (CPR) comprising:

a first region encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 1-19, 24-47, and 52-55; and a secondregion selected from the group consisting of: (i) a linker or atargeting domain encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 48-51; (ii) at least a portion of aprotein selected from the group consisting of; myelin oligodendrocyteglycoprotein (MOG), glutamic acid decarboxylase 2 (GAD65), myelinassociated glycoprotein (MAG), peripheral myelin protein 22 (PMP22),thyroid peroxidase (TPO), voltage-gated potassium channel (VGKC),proteolipid protein (PLP), acetylcholine receptor (AChR), tribblespseudokinase 2 (TRIB2), N-methyl-D-aspartate (NMDA)-type glutamatereceptor (GluR), glutamate decarboxylase 2 (GAD2), Armadillo repeatcontaining 9 (ARMC9), Cytochrome P450 Family 21 Subfamily A Member 2(CYP21A2), calcium sensing receptor (CASR), nuclear autoantigenic spermprotein (NASP), insulin, thyroid stimulating hormone receptor (TSHR),thyroperoxidase, asioglycoprotein receptor, Cytochrome P450 Family 2Subfamily D Member 6 (CYP2D6), lactoferrin (LF), tissuetrans-glutaminase (TTG), H/K ATP-ase, Factor XIII (F8),beta2-glycoprotein I (Beta2-GPI), erythrocyte I/I, B2 integrin (ITGB2),granulocyte-colony stimulating factor (G-CSF), glycoprotein (GP)IIb/IIa, collagen II (COLII), fibrinogen (FBG) βα, myeloperoxidase(MPO), cardiac myosin (CYO), proteinase 3 (PRTN3), trichohyalin (TCHH),bullous pemphigoid associated (BP), glycoprotein 1 (GPI), laminin-332(LM332), transglutaminase (TGM), type VII collagen (COLVII), P80 Coilin(COIL), Desmoglein I (DSG1), Desmoglein III (DSG3), SRY-Box 10 (SOX10),small nuclear ribonucleoprotein U1 subunit (70SNRNP70), S-antigen (SAG),and Collagen alpha-3(IV) chain (α3(IV)NC1 collagen); (iii) an antibodyor an antibody fragment selected from the group consisting of: 3F8, 8H9,Abagovomab, Abciximab, Abituzumab, Abrezekimab, Abrilumab, Actoxumab,Adalimumab, Adecatumumab, Atidortoxumab, Aducanumab, Afasevikumab,Afelimomab, Alacizumab pego, Alemtuzumab, Alirocumab, Altumomabpentetate, Amatuximab, Anatumomab mafenatox, Andecaliximab, Anetumabravtansine, Anifrolumab, Anrukinzumab, Apolizumab, Aprutumab ixadotin,Arcitumomab, Ascrinvacumab, Aselizumab, Atezolizumab, Atinumab,Atorolimumab, Avelumab, Azintuxizumab vedotin, Bapineuzumab,Basiliximab, Bavituximab, BCD-100, Bectumomab, Begelomab, Belantamabmafodotin, Belimumab, Bemarituzumab, Benralizumab, Berlimatoxumab,Bermekimab, Bersanlimab, Bertilimumab, Besilesomab, Bevacizumab,Bezlotoxumab, Biciromab, Bimagrumab, Bimekizumab, Birtamimab,Bivatuzumab mertansine, Bleselumab, Blinatumomab, Blontuvetmab,Blosozumab, Bococizumab, Brazikumab, Brentuximab vedotin, Briakinumab,Brodalumab, Brolucizumab, Brontictuzumab, Burosumab, Cabiralizumab,Camidanlumab tesirine, Camrelizumab, Canakinumab, Cantuzumab mertansine,Cantuzumab ravtansine, Caplacizumab, Capromab pendetide, Carlumab,Carotuximab, Catumaxomab, cBR96-doxorubicin immunoconjugate,Cedelizumab, Cemiplimab, Cergutuzumab amunaleukin, Certolizumab pegol,Cetrelimab, Cetuximab, Cibisatamab, Cirmtuzumab, Citatuzumab bogatox,Cixutumumab, Clazakizumab, Clenoliximab, Clivatuzumab tetraxetan,Codrituzumab, Cofetuzumab pelidotin, Coltuximab ravtansine, Conatumumab,Concizumab, Cosfroviximab, Crenezumab, Crizanlizumab, Crotedumab,CR6261, Cusatuzumab, Dacetuzumab, Daclizumab, Dalotuzumab, Dapirolizumabpegol, Daratumumab, Dectrekumab, Demcizumab, Denintuzumab mafodotin,Denosumab, Depatuxizumab mafodotin, Derlotuximab biotin, Detumomab,Dezamizumab, Dinutuximab, Diridavumab, Domagrozumab, Dorlimomab aritox,Dostarlimab, Drozitumab, DS-8201, Duligotuzumab, Dupilumab, Durvalumab,Dusigitumab, Duvortuxizumab, Ecromeximab, Eculizumab, Edobacomab,Edrecolomab, Efalizumab, Efungumab, Eldelumab, Elezanumab, Elgemtumab,Elotuzumab, Elsilimomab, Emactuzumab, Emapalumab, Emibetuzumab,Emicizumab, Enapotamab vedotin, Enavatuzumab, Enfortumab vedotin,Enlimomab pegol, Enoblituzumab, Enokizumab, Enoticumab, Ensituximab,Epitumomab cituxetan, Epratuzumab, Eptinezumab, Erenumab, Erlizumab,Ertumaxomab, Etaracizumab, Etigilimab, Etrolizumab, Evinacumab,Evolocumab, Exbivirumab, Fanolesomab, Faralimomab, Faricimab,Farletuzumab, Fasinumab, FBTA05, Felvizumab, Fezakinumab, Fibatuzumab,Ficlatuzumab, Figitumumab, Firivumab, Flanvotumab, Fletikumab,Flotetuzumab, Fontolizumab, Foralumab, Foravirumab, Fremanezumab,Fresolimumab, Frovocimab, Frunevetmab, Fulranumab, Futuximab,Galcanezumab, Galiximab, Gancotamab, Ganitumab, Gantenerumab,Gatipotuzumab, Gavilimomab, Gedivumab, Gemtuzumab ozogamicin,Gevokizumab, Gilvetmab, Gimsilumab, Girentuximab, Glembatumumab vedotin,Golimumab, Gomiliximab, Gosuranemab, Guselkumab, Ianalumab, Ibalizumab,IBI308, Ibritumomab tiuxetan, Icrucumab, Idarucizumab, Ifabotuzumab,Igovomab, Iladatuzumab vedotin, IMAB362, Imalumab, Imaprelimab,Imciromab, Imgatuzumab, Inclacumab, Indatuximab ravtansine, Indusatumabvedotin, Inebilizumab, Infliximab, Intetumumab, Inolimomab, Inotuzumabozogamicin, Ipilimumab, Iomab-B, Iratumumab, Isatuximab, Iscalimab,Istiratumab, Itolizumab, Ixekizumab, Keliximab, Labetuzumab,Lacnotuzumab, Ladiratuzumab vedotin, Lampalizumab, Lanadelumab,Landogrozumab, Laprituximab emtansine, Larcaviximab, Lebrikizumab,Lemalesomab, Lendalizumab, Lenvervimab, Lenzilumab, Lerdelimumab,Leronlimab, Lesofavumab, Letolizumab, Lexatumumab, Libivirumab,Lifastuzumab vedotin, Ligelizumab, Loncastuximab tesirine, Losatuxizumabvedotin, Lilotomab satetraxetan, Lintuzumab, Lirilumab, Lodelcizumab,Lokivetmab, Lorvotuzumab mertansine, Lucatumumab, Lulizumab pegol,Lumiliximab, Lumretuzumab, Lupartumab amadotin, Lutikizumab,Mapatumumab, Margetuximab, Marstacimab, Maslimomab, Mavrilimumab,Matuzumab, Mepolizumab, Metelimumab, Milatuzumab, Minretumomab,Mirikizumab, Mirvetuximab soravtansine, Mitumomab, Modotuximab,Mogamulizumab, Monalizumab, Morolimumab, Mosunetuzumab, Motavizumab,Moxetumomab pasudotox, Muromonab-CD3, Nacolomab tafenatox, Namilumab,Naptumomab estafenatox, Naratuximab emtansine, Narnatumab, Natahzumab,Navicixizumab, Navivumab, Naxitamab, Nebacumab, Necitumumab,Nemolizumab, NEOD001, Nerelimomab, Nesvacumab, Netakimab, Nimotuzumab,Nirsevimab, Nivolumab, Nofetumomab merpentan, Obiltoxaximab,Obinutuzumab, Ocaratuzumab, Ocrelizumab, Odulimomab, Ofatumumab,Olaratumab, Oleclumab, Olendalizumab, Olokizumab, Omalizumab,Omburtamab, OMS721, Onartuzumab, Ontuxizumab, Onvatilimab, Opicinumab,Oportuzumab monatox, Oregovomab, Orticumab, Otelixizumab, Otilimab,Otlertuzumab, Oxelumab, Ozanezumab, Ozoralizumab, Pagibaximab,Palivizumab, Pamrevlumab, Panitumumab, Pankomab, Panobacumab,Parsatuzumab, Pascolizumab, Pasotuxizumab, Pateclizumab, Patritumab,PDR001, Pembrolizumab, Pemtumomab, Perakizumab, Pertuzumab, Pexelizumab,Pidilizumab, Pinatuzumab vedotin, Pintumomab, Placulumab, Plozalizumab,Pogalizumab, Polatuzumab vedotin, Ponezumab, Porgaviximab, Prasinezumab,Prezalizumab, Priliximab, Pritoxaximab, Pritumumab, PRO 140, Quilizumab,Racotumomab, Radretumab, Rafivirumab, Ralpancizumab, Ramucirumab,Ranevetmab, Ranibizumab, Raxibacumab, Ravagalimab, Ravulizumab,Refanezumab, Regavirumab, Relatlimab, Remtolumab, Reslizumab,Rilotumumab, Rinucumab, Risankizumab, Rituximab, Rivabazumab pegol,Robatumumab, Rmab, Roledumab, Romilkimab, Romosozumab, Rontalizumab,Rosmantuzumab, Rovalpituzumab tesirine, Rovelizumab, Rozanolixizumab,Ruplizumab, SA237, Sacituzumab govitecan, Samalizumab, Samrotamabvedotin, Sarilumab, Satralizumab, Satumomab pendetide, Secukinumab,Selicrelumab, Seribantumab, Setoxaximab, Setrusumab, Sevirumab,Sibrotuzumab, SGN-CD19A, SHP647, Sifalimumab, Siltuximab, Simtuzumab,Siplizumab, Sirtratumab vedotin, Sirukumab, Sofituzumab vedotin,Solanezumab, Solitomab, Sonepcizumab, Sontuzumab, Spartalizumab,Stamulumab, Sulesomab, Suptavumab, Sutimlimab, Suvizumab, Suvratoxumab,Tabalumab, Tacatuzumab tetraxetan, Tadocizumab, Talacotuzumab,Talizumab, Tamtuvetmab, Tanezumab, Taplitumomab paptox, Tarextumab,Tavolimab, Tefibazumab, Telimomab aritox, Telisotuzumab vedotin,Tenatumomab, Teneliximab, Teplizumab, Tepoditamab, Teprotumumab,Tesidolumab, Tetulomab, Tezepelumab, TGN1412, Tibulizumab,Tildrakizumab, Tigatuzumab, Timigutuzumab, Timolumab, Tiragotumab,Tislelizumab, Tisotumab vedotin, TNX-650, Tocilizumab, Tomuzotuximab,Toralizumab, Tosatoxumab, Tositumomab, Tovetumab, Tralokinumab,Trastuzumab, Trastuzumab emtansine, TRBS07, Tregalizumab, Tremelimumab,Trevogrumab, Tucotuzumab celmoleukin, Tuvirumab, Ublituximab,Ulocuplumab, Urelumab, Urtoxazumab, Ustekinumab, Utomilumab,Vadastuximab talirine, Vanalimab, Vandortuzumab vedotin, Vantictumab,Vanucizumab, Vapaliximab, Varisacumab, Varlilumab, Vatelizumab,Vedolizumab, Veltuzumab, Vepalimomab, Vesencumab, Visilizumab,Vobarilizumab, Volociximab, Vonlerolizumab, Vopratelimab, Vorsetuzumabmafodotin, Votumumab, Vunakizumab, Xentuzumab, XMAB-5574, Zalutumumab,Zanolimumab, Zatuximab, Zenocutuzumab, Ziralimumab, Zolbetuximab,Zolimomab aritox; and (iv) a major histocompatibility complex (MHC)class 1 receptor or a major histocompatibility complex (MHC) class 2receptor, wherein the MHC class 1 receptor is bound to a peptide derivedfrom a tumor antigen, a neoantigen, or an autoantigen or the MHC class 2receptor is bound to a peptide derived from a tumor antigen, aneoantigen, or an autoantigen.

In some embodiments, the chimeric platelet receptor binds at least oneantigen. In some embodiments, the chimeric platelet receptor binds atissue in the body of a subject. In some embodiments, the chimericplatelet receptor inhibits platelet activation. In some embodiments, thechimeric platelet receptor is an ITIM-containing receptor. In someembodiments, the chimeric platelet receptor stimulates plateletactivation. In some embodiments, the chimeric platelet receptor is anITAM-containing receptor. In some embodiments the chimeric plateletreceptor binds to at least one antigen that is an endogenous antigen,for example is an endogenous host or subject antigen. By an endogenoushost or subject antigen we include the meaning that the antigen ispresent within a host or subject into which the chimeric plateletreceptor is administered or into which cells such as megakaryocytes orplatelets that comprise the CPR of the invention are administered. Forexample the antigen may be an antigen that is present on tissue, or on aparticular subset of tissue, or in plasma or blood of a subject, forexample a human subject. The antigen may be an antigen that is expressedat abnormal levels, for example at abnormally high levels, on a tissuethat does not normally express a high level of the antigen, or that doesnot normally express the antigen at all.

In some embodiments, the chimeric platelet receptor is not, or does notcomprise, a GPCR or a protease-activated receptor.

The invention also provides a nucleic acid encoding the chimericplatelet receptor of the invention. In preferred instances, the chimericplatelet receptor is not a naturally occurring receptor, and so thenucleic acid encoding said receptor is not a naturally occurring nucleicacid. In some embodiments the nucleic acid encodes the CPR of theinvention and also comprises a heterologous nucleic acid sequence. Insome instances the nucleic acid is operatively linked to an expressioncontrol sequence. Expression control sequences are considered to includecomponent such as enhancers and promoters. In one embodiment the nucleicacid of the invention comprises a heterologous promoter. In the same ordifferent embodiment the nucleic acid of the invention comprises aheterologous enhancer sequence.In some embodiments the nucleic acid is DNA. In some embodiments thenucleic acid is RNA for example is an mRNA. In some embodiments thenucleic acid comprises a megakaryocyte-specific promoter or aplatelet-specific promoter. The terms megakaryocyte-specific promoterand platelet-specific promoter are used synonymously. The skilled personwill understand what is meant by the terms megakaryocyte-specificpromoter and platelet-specific promoter.The invention also provides a vector that comprises a nucleic acid thatencodes the CPR By vector we include the meaning of plasmid. In someembodiments the vector also comprises a heterologous nucleic acid. Insome embodiments the vector comprises a megakaryocyte-specific promoter.In some embodiments the vector comprises a platelet-specific promoter.The invention also provides a viral particle, or viral vector,comprising any one or more of the nucleic acids of the invention.The invention also provides a nucleic acid encoding a cargo protein orpeptide which comprises sequences suitable for driving expression in amegakaryocyte and/or platelet. For example, in some embodiments thenucleic acid encoding the cargo protein, cargo peptide or cargo RNA isoperatively linked to a heterologous expression control sequence such asa promoter. In some embodiments the nucleic acid encodes a cargo proteinor peptide and also comprises a megakaryocyte specific promoter or aplatelet specific promoter. In some embodiments the nucleic acid encodesa cargo protein or peptide and comprises a heterologous sequence, suchas a megakaryocyte specific promoter or a platelet specific promoter.

Various embodiments of the invention described herein provide atherapeutic delivery system comprising: an engineered plateletpresenting the chimeric platelet receptor previously described; and atleast one therapeutic agent selected from the group consisting of: atoxin, a protein, a small molecule drug, and a nucleic acid packagedwithin a vesicle inside the platelet.

In some embodiments, the engineered platelet is produced from an iPSCprogenitor. In some embodiments, the nucleic acid is a mRNA, a miRNA,shRNA, and a clustered regularly interspaced short palindromic repeats(CRISPR) sequence. In some embodiments, the protein is selected from thegroup consisting of an antibody, an enzyme, a cytokine, and a CRISPRassociated protein 9 (Cas9). In an aspect, the enzyme is a nuclease.

In some embodiments, the nuclease is a transcription activator-likeeffector nuclease (TALEN). In some embodiments, the antibody binds atarget such as, but not limited to, a tumor antigen or a neoantigen. Insome embodiments, the therapeutic agent is released from the plateletfollowing activation of the platelet by an antigen recognized by thechimeric platelet receptor.

Various methods are provided for delivering a cargo to a subject in needthereof. As described herein, the cargo may be a therapeutic drug or atoxin. The cargo may be a protein or peptide, or may be a nucleic acidsuch as a therapeutic RNA or an mRNA. Preferences for the cargo are asdescribed elsewhere herein. The invention provides a method ofdelivering a cargo comprising administering an effective amount of anyone or more of an engineered megakaryocyte, engineered platelet, and/orCPR according to any of the preceding claims. The invention alsoprovides a therapeutic delivery system. The invention also provides anon-therapeutic delivery system. The invention also provides a method oftargeted cargo delivery to a target tissue or site in the body whereinthe method comprises administering an effective amount of any one ormore of an engineered megakaryocyte, engineered platelet, and/or CPRaccording to any of the preceding claims wherein the targeting domain ofthe CPR binds to the target tissue or site in the body.Various embodiments of the invention described herein provide a methodof treating a disease, disorder, or condition in a subject, the methodcomprising: administering to the subject the previously describedtherapeutic delivery system, wherein the chimeric receptor is specificto an antigen associated with the disease, disorder, or condition.

In some embodiments, the disease, disorder, or condition may be, but isnot limited to, a cancer, an autoimmunity, and an infection. In someembodiments, the cancer is selected from the group consisting of: Acutegranulocytic leukemia, Acute lymphocytic leukemia, Acute myelogenousleukemia, Adenocarcinoma, Adenosarcoma, Adrenal cancer, Adrenocorticalcarcinoma, Anal cancer, Anaplastic astrocytoma, Angiosarcoma, Appendixcancer, Astrocytoma, Basal cell carcinoma, B-Cell lymphoma), Bile ductcancer, Bladder cancer, Bone cancer, Bowel cancer, Brain cancer, Brainstem glioma, Brain tumor, Breast cancer, Carcinoid tumors, Cervicalcancer, Cholangiocarcinoma, Chondrosarcoma, Chronic lymphocyticleukemia, Chronic myelogenous leukemia, Colon cancer, Colorectal cancer,Craniopharyngioma, Cutaneous lymphoma, Cutaneous melanoma, Diffuseastrocytoma, Ductal carcinoma in situ, Endometrial cancer, Ependymoma,Epithelioid sarcoma, Esophageal cancer, Ewing sarcoma, Extrahepatic bileduct cancer, Eye cancer, Fallopian tube cancer, Fibrosarcoma,Gallbladder cancer, Gastric cancer, Gastrointestinal cancer,Gastrointestinal carcinoid cancer, Gastrointestinal stromal tumors,General, Germ cell tumor, Glioblastoma multiforme, Glioma, Hairy cellleukemia, Head and neck cancer, Hemangioendothelioma, Hodgkin lymphoma,Hodgkin's disease, Hodgkin's lymphoma, Hypopharyngeal cancer,Infiltrating ductal carcinoma, Infiltrating lobular carcinoma,Inflammatory breast cancer, Intestinal Cancer, Intrahepatic bile ductcancer, Invasive/infiltrating breast cancer, Islet cell cancer, Jawcancer, Kaposi sarcoma, Kidney cancer, Laryngeal cancer, Leiomyosarcoma,Leptomeningeal metastases, Leukemia, Lip cancer, Liposarcoma, Livercancer, Lobular carcinoma in situ, Low-grade astrocytoma, Lung cancer,Lymph node cancer, Lymphoma, Male breast cancer, Medullary carcinoma,Medulloblastoma, Melanoma, Meningioma, Merkel cell carcinoma,Mesenchymal chondrosarcoma, Mesenchymous, Mesothelioma, Metastaticbreast cancer, Metastatic melanoma, Metastatic squamous neck cancer,Mixed gliomas, Mouth cancer, Mucinous carcinoma, Mucosal melanoma,Multiple myeloma, Nasal cavity cancer, Nasopharyngeal cancer, Neckcancer, Neuroblastoma, Neuroendocrine tumors, Non-Hodgkin lymphoma,Non-Hodgkin's lymphoma, Non-small cell lung cancer, Oat cell cancer,Ocular cancer, Ocular melanoma, Oligodendroglioma, Oral cancer, Oralcavity cancer, Oropharyngeal cancer, Osteogenic sarcoma, Osteosarcoma,Ovarian cancer, Ovarian epithelial cancer, Ovarian germ cell tumor,Ovarian primary peritoneal carcinoma, Ovarian sex cord stromal tumor,Paget's disease, Pancreatic cancer, Papillary carcinoma, Paranasal sinuscancer, Parathyroid cancer, Pelvic cancer, Penile cancer, Peripheralnerve cancer, Peritoneal cancer, Pharyngeal cancer, Pheochromocytoma,Pilocytic astrocytoma, Pineal region tumor, Pineoblastoma, Pituitarygland cancer, Primary central nervous system lymphoma, Prostate cancer,Rectal cancer, Renal cell cancer, Renal pelvis cancer, Rhabdomyosarcoma,Salivary gland cancer, Sarcoma, Sarcoma, bone, Sarcoma, soft tissue,Sarcoma, uterine, Sinus cancer, Skin cancer, Small cell lung cancer,Small intestine cancer, Soft tissue sarcoma, Spinal cancer, Spinalcolumn cancer, Spinal cord cancer, Spinal tumor, Squamous cellcarcinoma, Stomach cancer, Synovial sarcoma, T-cell lymphoma),Testicular cancer, Throat cancer, Thymoma/thymic carcinoma, Thyroidcancer, Tongue cancer, Tonsil cancer, Transitional cell cancer,Transitional cell cancer, Transitional cell cancer, Triple-negativebreast cancer, Tubal cancer, Tubular carcinoma, Ureteral cancer,Ureteral cancer, Urethral cancer, Uterine adenocarcinoma, Uterinecancer, Uterine sarcoma, Vaginal cancer, and Vulvar cancer.

In some embodiments, the method further comprises incubating theengineered platelet with the at least one therapeutic agent such as, butnot limited to, a toxin, a protein, and a small molecule drug to producethe therapeutic delivery system. In some embodiments, the nucleic acidmay be, but is not limited to, a mRNA, a miRNA, shRNA, and a clusteredregularly interspaced short palindromic repeats (CRISPR) sequence. Insome embodiments, the protein may be, but is not limited to, anantibody, an enzyme, and a CRISPR associated protein 9 (Cas9). In someembodiments, the enzyme is a nuclease. In some embodiments, the nucleaseis a transcription activator-like effector nuclease (TALEN). In someembodiments, incubating occurs prior to administering. In someembodiments, the disease, disorder, or condition is an autoimmunity suchas, but not limited to, Autoimmune disseminated encephalomyelitis,Autoimmune inner ear disease, Batten disease/Neuronal CeroidLipofuscinoses, Chronic inflammatory demyelinating polyneuropathy,Encephalitis lethargica, Anti-basal ganglia, Guillain-Barré syndrome,Hashimoto's Encephalopathy, Anti-TPO, Isaac's syndrome/acquiredneuromyotonia, Miller Fisher syndrome Morvan's syndrome, Multiplesclerosis, Myasthenia gravis, Narcolepsy PANDAS, Rasmussen'sencephalitis, Stiff-person syndrome, Vogt-Koyanagi-Harada syndrome,Addison's disease, Autoimmune hypoparathyroidism, Autoimmunehypophysitis, Autoimmune oophoritis, Autoimmune orchitis, Autoimmunepolyglandular syndrome I (APECED), Autoimmune polyglandular syndrome II,Autoimmune polyglandular syndrome III, Diabetes mellitus, type 1,Graves' disease, Hashimoto's autoimmune thyroiditis,Immunodysregulation, polyendocrinopathy, enteropathy, X-linked,Autoimmune hepatitis type 1, Autoimmune hepatitis type 2, Autoimmunepancreatitis, Coeliac disease, Crohn's disease, Perniciousanemia/atrophic gastritis, Primary biliary cirrhosis, Primary sclerosingcholangitis, Ulcerative colitis, Acquired hemophilia A, Antiphospholipidsyndrome, Autoimmune hemolytic anemia, Autoimmune lymphoproliferativesyndrome, Autoimmune neutropenia, Evans syndrome, Felty's syndrome,Immune thrombocytopenic purpura, Polymyositis/dermatomyositis, Relapsingpolychondritis, Rheumatoid arthritis, Still's disease, Alopecia areata,Bullous pemphigoid, Cicatricial pemphigoid, Dermatitis herpetiformis,Discoid lupus erythematosus, Epidermolysis bullosa acquisita, Linearmorphea, Pemphigus foliaceus, Pemphigus vulgaris, Vitiligo, Behçetdisease, Churg-Strauss syndrome, Cogan's syndrome, CREST syndrome,Anti-fibrillarin, Essential mixed cryoglobulinemia, Mixed connectivetissue disease, POEMS syndrome, Scleroderma, Sjögren's syndrome,Systemic lupus erythematosus, Erythema elevatum diutinum, Kawasakidisease, Microscopic polyangiitis, Polyarteritis nodosa, Rheumaticfever, Takayasu arteritis Temporal arteritis, Wegener's granulomatosis,HLA-B27-associated acute anterior uveitis, Sympathetic ophthalmia, andGoodpasture's disease.

The invention also provides non-therapeutic methods of delivering acargo to a subject in need or desirous thereof. For example theinvention provides a non-therapeutic method of delivering cargo to asubject in need thereof. In such non-therapeutic methods, the cargo maybe a cosmetic agent. In some embodiments the invention provides anon-therapeutic method of targeted delivery of a cargo to a subject inneed thereof.

Various embodiments of the invention described herein provide anengineered platelet produced from a megakaryocyte comprising a mutationin the nucleic acid sequence resulting in disruption of a vesiclebiogenesis pathway or a vesicle release pathway in the platelet,expression of a toxin, or expression of for example a cargo that is aprotein or peptide or a RNA for example an mRNA, for example atherapeutic agent or imaging agent, or deletion of a platelet receptor,mediator, or signal transduction protein compared to a platelet producedfrom a megakaryocyte without the mutation.

In some embodiments, the megakaryocyte is differentiated from an iPSCprogenitor or the megakaryocyte is immortalized. In some embodiments,the mutation occurs in a gene encoding a component of the vesiclebiogenesis pathway or a vesicle release pathway of the engineeredplatelet such as, but not limited to, α-granules, dense granules, andlarge dense-core vesicle. In some embodiments, the mutation is aninsertion of a gene encoding a major histocompatibility complex (MHC)molecule. In some embodiments, the deletion is of at least one gene suchas, but not limited to, RAB27a (RAS oncogene), HPS (haptoglobin) genes,integrin AIIbB3, GP1b-IX-V (Glycoprotein Ib complexed with glycoproteinIX), Par1 (protease activated receptor 1), Par4 (protease activatedreceptor 4), P2Y1 (purinergic receptor P2Y1), P2Y12 (purinergic receptorP2Y12), IP (PGI2R or prostaglandin 12 receptor), TP (TxA2R orThromboxane A2 Receptor), TLR (toll-like receptor), GPVI, a2B1 (type 1collagen receptor), GPIIbIIIA (Glycoprotein IIb Platelet Subunit Alpha),CLEC-2 (C-type lectinlike receptor 2), MyD88 (Myeloid DifferentiationPrimary Response 88), Galphaq (G-protein alpha pathway q), LIMK1 (LIMDomain Kinase 1), vWF (von Willebrand), Fibrinogen, PDGF (plateletderived growth factor), VEGF (vascular endothelial growth factor),Factor V, Factor VIII, Factor XI, Factor XIII, PF4 (platelet factor 4),NAP2 (Nucleosome Assembly Protein 2), Prothrombin, High Molecular WeightKininogens, Plasminogen activator inhibitor 1, a2-antiplasmin,plasminogen, P-Selectin, CXCL4 (C-X-C motif chemokine ligand 4), CXCL7(C-X-C motif chemokine ligand 7), FGF (fibroblast growth factor), EGF(elongation growth factor), HGF (hepatocyte growth factor), IGF(insulin-like growth factor), Angipoetin, Thromboxane synthase, PAF(platelet activating factor), cPLA2a, Thromospondin, CD40L, SgIII(Secretogranin III), Endostatin, TGF-β (transforming growth factorbeta), Talin1, Kindlins, and Anoctamin 6.

In some embodiments, the mutation is a deletion which is a knock-out ofa gene encoding a pro-thrombotic factor. In some embodiments, the geneis a 02 microglobulin gene, wherein the deletion results in endogenousMHC class 1 disruption and the generation of a non-immunogenic platelet.In some embodiments, the mutation reduces the thrombogenic potential ofthe engineered platelet compared to a platelet produced from amegakaryocyte without the mutation.

Various embodiments of the invention described herein provide a methodof reducing activity in the immune system of a subject, the methodcomprising: administering to the subject an engineered plateletpresenting at least one receptor expressing a major histocompatibilitycomplex (MHC) molecule bound to a peptide derived from a tumor antigen,a neoantigen, or an autoantigen.

In some embodiments, the receptor expresses a MHC class I molecule. Insome embodiments, the receptor expresses a MHC class H molecule. In someembodiments, wherein the MHC molecule stimulates an immune response toan antigen. In some embodiments, the antigen is associated with at leastone disease, disorder, or condition selected from the group consistingof: a cancer, an autoimmunity, and an infection.

Various embodiments of the invention described herein provide a methodof in vitro production of platelets, the method comprising: transfectinga plurality of induced pluripotent stem cell (iPSC) progenitors with anexpression system, wherein the expression system is induced by an agentnot found in an iPSC; establishing a megakaryocyte progenitor cell lineby contacting the expression system with the agent to expandmegakaryocytes; engineering the megakaryocyte to have at least onemutation such as, but not limited to, insertion of a nucleic sequenceencoding a chimeric platelet receptor previously described, insertion ofa nucleic acid sequence encoding a toxin, or for example encoding acargo that is a protein or peptide or a RNA for example an mRNA, forexample a therapeutic agent or imaging agent, and deletion of a nucleicacid sequence encoding a platelet receptor; and removing the agent fromthe expression system to induce differentiation of the megakaryocytesinto platelets.

In some embodiments, the mutation results in platelets with lessimmunogenicity compared to platelets from human donors. In someembodiments, the platelet does not function analogously to plateletsderived from a human donor. In some embodiments, the deletion preventstoxin release or prevents cargo release in response to plateletactivation signals. In some embodiments, the toxin or cargo is attachedto an α-granule localization signal. In some embodiments, the α-granulelocalization signal. In some embodiments, the method of plateletproduction further comprising contacting the platelets with at least oneof a cargo for example a cargo that is a protein or peptide or a RNA forexample an mRNA, for example a therapeutic agent or imaging agent or asmall molecule; a toxin; and a small molecule drug under conditions tofacilitate absorption by the platelet. In some embodiments, theexpression system further comprises a platelet-specific promoter.

Various embodiments of the invention described herein provide a methodof in vivo gene editing or gene therapy in a subject, the methodcomprising: administering to the subject an engineered plateletcomprising a chimeric platelet receptor described herein specific to atissue to be edited, wherein the engineered platelet is cloaking anadenovirus loaded with genome engineering machinery; and releasing thegenome machinery at the tissue. In some embodiments, the genomemachinery is a CRISPR/Cas gene editing system.

Various embodiments of the invention described herein provide a use ofthe therapeutic delivery system previously described, wherein thechimeric receptor is specific to an antigen associated with the disease,disorder, or condition in treating a disease, disorder, or condition ina subject. In some embodiments of the use described herein, the disease,disorder, or condition is selected from the group consisting of: acancer, an autoimmunity, and an infection.

In some embodiments of the use described herein, the cancer may be, butis not limited to, Acute granulocytic leukemia, Acute lymphocyticleukemia, Acute myelogenous leukemia, Adenocarcinoma, Adenosarcoma,Adrenal cancer, Adrenocortical carcinoma, Anal cancer, Anaplasticastrocytoma, Angiosarcoma, Appendix cancer, Astrocytoma, Basal cellcarcinoma, B-Cell lymphoma), Bile duct cancer, Bladder cancer, Bonecancer, Bowel cancer, Brain cancer, Brain stem glioma, Brain tumor,Breast cancer, Carcinoid tumors, Cervical cancer, Cholangiocarcinoma,Chondrosarcoma, Chronic lymphocytic leukemia, Chronic myelogenousleukemia, Colon cancer, Colorectal cancer, Craniopharyngioma, Cutaneouslymphoma, Cutaneous melanoma, Diffuse astrocytoma, Ductal carcinoma insitu, Endometrial cancer, Ependymoma, Epithelioid sarcoma, Esophagealcancer, Ewing sarcoma, Extrahepatic bile duct cancer, Eye cancer,Fallopian tube cancer, Fibrosarcoma, Gallbladder cancer, Gastric cancer,Gastrointestinal cancer, Gastrointestinal carcinoid cancer,Gastrointestinal stromal tumors, General, Germ cell tumor, Glioblastomamultiforme, Glioma, Hairy cell leukemia, Head and neck cancer,Hemangioendothelioma, Hodgkin lymphoma, Hodgkin's disease, Hodgkin'slymphoma, Hypopharyngeal cancer, Infiltrating ductal carcinoma,Infiltrating lobular carcinoma, Inflammatory breast cancer, IntestinalCancer, Intrahepatic bile duct cancer, Invasive/infiltrating breastcancer, Islet cell cancer, Jaw cancer, Kaposi sarcoma, Kidney cancer,Laryngeal cancer, Leiomyosarcoma, Leptomeningeal metastases, Leukemia,Lip cancer, Liposarcoma, Liver cancer, Lobular carcinoma in situ,Low-grade astrocytoma, Lung cancer, Lymph node cancer, Lymphoma. Malebreast cancer, Medullary carcinoma, Medulloblastoma, Melanoma,Meningioma, Merkel cell carcinoma, Mesenchymal chondrosarcoma,Mesenchymous, Mesothelioma, Metastatic breast cancer, Metastaticmelanoma, Metastatic squamous neck cancer, Mixed gliomas, Mouth cancer,Mucinous carcinoma, Mucosal melanoma, Multiple myeloma, Nasal cavitycancer, Nasopharyngeal cancer, Neck cancer, Neuroblastoma,Neuroendocrine tumors, Non-Hodgkin lymphoma, Non-Hodgkin's lymphoma,Non-small cell lung cancer, Oat cell cancer, Ocular cancer, Ocularmelanoma, Oligodendroglioma, Oral cancer, Oral cavity cancer,Oropharyngeal cancer, Osteogenic sarcoma, Osteosarcoma, Ovarian cancer,Ovarian epithelial cancer, Ovarian germ cell tumor, Ovarian primaryperitoneal carcinoma, Ovarian sex cord stromal tumor, Paget's disease,Pancreatic cancer, Papillary carcinoma, Paranasal sinus cancer,Parathyroid cancer, Pelvic cancer, Penile cancer, Peripheral nervecancer, Peritoneal cancer, Pharyngeal cancer, Pheochromocytoma,Pilocytic astrocytoma, Pineal region tumor, Pineoblastoma, Pituitarygland cancer, Primary central nervous system lymphoma, Prostate cancer,Rectal cancer, Renal cell cancer, Renal pelvis cancer, Rhabdomyosarcoma,Salivary gland cancer, Sarcoma, Sarcoma, bone, Sarcoma, soft tissue,Sarcoma, uterine, Sinus cancer, Skin cancer, Small cell lung cancer,Small intestine cancer, Soft tissue sarcoma, Spinal cancer, Spinalcolumn cancer, Spinal cord cancer, Spinal tumor, Squamous cellcarcinoma, Stomach cancer, Synovial sarcoma, T-cell lymphoma),Testicular cancer, Throat cancer, Thymoma/thymic carcinoma, Thyroidcancer, Tongue cancer, Tonsil cancer, Transitional cell cancer,Transitional cell cancer, Transitional cell cancer, Triple-negativebreast cancer, Tubal cancer, Tubular carcinoma, Ureteral cancer,Ureteral cancer, Urethral cancer, Uterine adenocarcinoma, Uterinecancer, Uterine sarcoma, Vaginal cancer, and Vulvar cancer.

In some embodiments of the use described herein, the disease, disorder,or condition is an autoimmunity such as, but not limited to, Autoimmunedisseminated encephalomyelitis, Autoimmune inner ear disease, Battendisease/Neuronal Ceroid Lipofuscinoses, Chronic inflammatorydemyelinating polyneuropathy, Encephalitis lethargica, Anti-basalganglia, Guillain-Barré syndrome, Hashimoto's Encephalopathy, Anti-TPO,Isaac's syndrome/acquired neuromyotonia, Miller Fisher syndrome Morvan'ssyndrome, Multiple sclerosis, Myasthenia gravis, Narcolepsy PANDAS,Rasmussen's encephalitis, Stiff-person syndrome, Vogt-Koyanagi-Haradasyndrome, Addison's disease, Autoimmune hypoparathyroidism, Autoimmunehypophysitis, Autoimmune oophoritis, Autoimmune orchitis, Autoimmunepolyglandular syndrome I (APECED), Autoimmune polyglandular syndrome II,Autoimmune polyglandular syndrome III, Diabetes mellitus, type 1,Graves' disease, Hashimoto's autoimmune thyroiditis,Immunodysregulation, polyendocrinopathy, enteropathy, X-linked,Autoimmune hepatitis type 1, Autoimmune hepatitis type 2, Autoimmunepancreatitis, Coeliac disease, Crohn's disease, Perniciousanemia/atrophic gastritis, Primary biliary cirrhosis, Primary sclerosingcholangitis, Ulcerative colitis, Acquired hemophilia A, Antiphospholipidsyndrome, Autoimmune hemolytic anemia, Autoimmune lymphoproliferativesyndrome, Autoimmune neutropenia, Evans syndrome, Felty's syndrome,Immune thrombocytopenic purpura, Polymyositis/dermatomyositis, Relapsingpolychondritis, Rheumatoid arthritis, Still's disease, Alopecia areata,Bullous pemphigoid, Cicatricial pemphigoid, Dermatitis herpetiformis,Discoid lupus erythematosus, Epidermolysis bullosa acquisita, Linearmorphea, Pemphigus foliaceus, Pemphigus vulgaris, Vitiligo, Behçetdisease, Churg-Strauss syndrome, Cogan's syndrome, CREST syndrome,Anti-fibrillarin, Essential mixed cryoglobulinemia, Mixed connectivetissue disease, POEMS syndrome, Scleroderma, Sjögren's syndrome,Systemic lupus erythematosus, Erythema elevatum diutinum, Kawasakidisease, Microscopic polyangiitis, Polyarteritis nodosa, Rheumaticfever, Takayasu arteritis Temporal arteritis, Wegener's granulomatosis,HLA-B27-associated acute anterior uveitis, Sympathetic ophthalmia, andGoodpasture's disease.

Various embodiments of the invention herein provide a therapeuticdelivery system comprising: (a) an engineered platelet presenting thechimeric platelet receptor, wherein the engineered platelet has beenproduced through genetic modification of a progenitor megakaryocyte tobe non-thrombogenic and non-immunogenic; and (b) at least onetherapeutic agent selected from the group consisting of: a cargo asdefined herein, a toxin, a protein, a small molecule drug, and a nucleicacid packaged within a vesicle inside the platelet, i) wherein thetherapeutic agent is the nucleic acid or the protein, loading occursthrough expression in a progenitor megakaryocyte, or ii) wherein thetherapeutic agent is loaded by incubation of the engineered plateletwith the therapeutic agent.

DETAILED DESCRIPTION OF THE INVENTION

I. Introduction

Genetically engineered platelets generated outside of the body may betargeted to respond to specific proteins expressed on a target tissue orcell (e.g., on cancer) and release a therapeutic agent (e.g., a smallmolecule drug, antibody, or a nucleic acid encoding a protein). Theseengineered platelets would become targeted drug delivery devices. Unlikewhen transplanting other tissues, platelets require limited matching toa recipient's immune system and thus, the engineered platelets couldfunction “off the shelf,” without having to be matched to a specificpatient (which is a major problem with current cell therapies).

The engineered platelets described herein may be generated outside thebody from megakaryocytes. As the megakaryocyte is maintained in cultureoutside of the body, it can be extensively edited at the genome level(e.g. by CRISPR/Cas9) without fear of oncogenic transformation in thepatient, which is not possible with other competing cell therapyproducts. The engineered platelets would have a lifespan in the body of7-10 days, with little to no potential for continued reproduction, thuslittle to no chance of forming a tumour itself.

Platelets can be frozen and stored for extended period of time resultingin an extended shelf life, and with currently available technology,engineered platelets could be produced, stored, transported andadministered to patients without issue due to their lack ofimmunogenicity.

Engineered platelets could be stripped of all thrombogenic potential bygenome editing of megakaryocytes in vitro to alleviate potentialthrombotic safety concerns. An engineered platelet, also called aSYNLET™ therapeutic delivery system, may act as a blank template,effectively functioning as a scaffold, having the capacity to storecargo internally in vesicles, and genetic alterations of megakaryocytesallow the engineered platelets to respond to specific antigens orsignals. Platelets contain a variety of signaling pathways, thereforeengineered inputs could be positive or negative, allowing the engineeredplatelets to integrate and compute a variety of stimuli before makingthe decision to activate. Interestingly, platelets also have thecapacity to transfer RNA to nearby cells providing the potential togenetically manipulate target cells. For example, a CRISPR/Cas9 systemmay be delivered to a target cell by an engineered platelet.Additionally, signaling pathways within platelets can trigger the denovo translation of proteins, so these could be harnessed to deliverpotentially toxic proteins locally to target locations.

In contrast to chimeric antigen receptor T (CAR-T) cells, the presentinvention provides an engineered platelet that is a universal productwhich does not require a match to a patient before administration.Further, platelet production in vitro from progenitors described herein,removes the need to continuously produce virus and edit cells. Due tothe short life span of the engineered platelets described herein, safetyconcerns are limited as compared to current gene editing therapeutics.For example, gene editing and genome stability is less of a concern thanwith CAR-T cells because platelets are enucleate and therefore thecomplexity of the platelet therapy is not limited by the efficiency ofediting or culture time limits. Additionally, due to their smaller size,the engineered platelets may provide better access to solid tumors thanCAR-T cells.

Enucleated red blood cells, such as those commercially available fromRubius Therapeutics, Inc., have also been contemplated in the art fordelivering therapeutic agents. In contrast to red blood cells, theengineered platelets described herein are highly metabolically activeand include signaling systems that can be re-engineered. In fact, moretargeted uses are possible with the engineered platelets compared to redblood cells. Vesicle degranulation of the platelets also allows for“hiding” of protein until the desired target is engaged, which is notpossible with enucleated red blood cells because the biotherapeuticproteins are generally expressed on the surface of the cell.Accordingly, in one embodiment binding of the targeting domain of thechimeric platelet receptor to the target or antigen results indegranulation of a platelet that comprises the chimeric plateletreceptor.

The engineered platelets described herein are smaller than red bloodcells likely resulting in better biodistribution.

II. Gene Nomenclature

Gene symbols are used herein, along with ENSEMBL Gene IDs, to refer togenes from humans. Unless otherwise noted, the gene name and ENSEMBLGene (ENSG) IDs corresponding to each gene symbol are shown in Table 1.The unique identifiers for each ENSEMBL entry has been modified toremove the first five leading zeros (0) of the identifier after the ENSGlabel.

TABLE 1 ENSEMBL identifiers for genes Given Symbol Given Name ENSGRAB27A RAS oncogene 069974 HP haptoglobin 257017 integrin AIIbB3 orITGB3 glycoprotein beta III platelet subunit alpha 2 259207 GP1b-IX-VGlycoprotein Ib complexed with glycoprotein IX N/A Par1 or NR1I2protease activated receptor 1 144852 Par4 or F2RL3 protease activatedreceptor 4 127533 P2Y1 or P2RY1 purinergic receptor P2Y1 169860 P2Y12 orP2RY12 purinergic receptor P2Y12 169313 IP or PTGIR PGI2R orprostaglandin I2 receptor 160013 TP or TBXA2R TxA2R or Thromboxane A2Receptor 006638 TLR1 toll-like receptor 1 174125 TLR2 toll-like receptor2 137462 TLR3 toll-like receptor 3 164342 TLR4 toll-like receptor 4136869 TLR5 toll-like receptor 5 187554 TLR6 toll-like receptor 6 174130TLR7 toll-like receptor 7 196664 TLR8 toll-like receptor 8 101916 TLR9toll-like receptor 9 239732 TLR10 toll-like receptor 10 174123 GPVI orGP6 glycoprotein VI 274050, 275931, 275633, 274566, 278316, 088053,278670, 277439, 276211, 276065 a2B1 or ITGB1 type 1 collagen receptor150093 GPIIbIIIA or ITGA2B Glycoprotein IIb Platelet Subunit Alpha005961 CLEC-2 or CLEC1B C-type lectinlike receptor 2 165682 MyD88 orMYD88 Myeloid Differentiation Primary Response 88 172936 Galphaq or GNAQG-protein alpha pathway q 156052 LIMK1 or LIMK1 LIM Domain Kinase 1106683 vWF or VWF von Willebrand 110799 FGA Fibrinogen 171560 FGBFibrinogen 171564 FGG Fibrinogen 171557 PDGFA platelet derived growthfactor A 197461 PDGFB platelet derived growth factor B 100311 PDGFCplatelet derived growth factor C 145431 PDGFD platelet derived growthfactor D 170962 VEGF or VEGFA vascular endothelial growth factor 112715F5 Factor V 198734 F8 Factor VIII 185010 F11 Factor XI 088926 F13A1Factor XIII A 124491 F13B Factor XIII B 143278 CXCL4 or PF4 C-X-C motifchemokine ligand 4 or platelet factor 4 163737, 163737 NAP2 or PPBPNucleosome Assembly Protein 2 163736 F2 Prothrombin 180210 PAI-1 orSERPINE1 Plasminogen activator inhibitor 1 106366 SERPIN or SERPINF1a2-antiplasmin 282307, 132386 PLG plasminogen 122194 SELP P-Selectin174175 CXCL7 or PPBP C-X-C motif chemokine ligand 7 163736 FGF1fibroblast growth factor 1 113578 FGF2 fibroblast growth factor 2 138685FGF3 fibroblast growth factor 3 186895 FGF4 fibroblast growth factor 4075388 FGF5 fibroblast growth factor 5 138675 FGF6 fibroblast growthfactor 6 111241 FGF7 fibroblast growth factor 7 140285 FGF8 fibroblastgrowth factor 8 107831 FGF9 fibroblast growth factor 9 102678 FGF10fibroblast growth factor 10 070193 FGF11 fibroblast growth factor 11161958 FGF12 fibroblast growth factor 12 283903 FGF13 fibroblast growthfactor 13 114279 FGF14 fibroblast growth factor 14 129682 FGF15fibroblast growth factor 15 102466 FGF16 fibroblast growth factor 16196468 FGF17 fibroblast growth factor 17 158815 FGF18 fibroblast growthfactor 18 156427 FGF19 fibroblast growth factor 19 162344 FGF20fibroblast growth factor 20 078579 FGF21 fibroblast growth factor 21105550 FGF22 fibroblast growth factor 22 070388 FGF23 fibroblast growthfactor 23 118972 EGF elongation growth factor 138798 HGF hepatocytegrowth factor 019991 IGF or IGF1 insulin-like growth factor 017427 ANGPTor ANGPT1 Angiopoetin 154188 TBXAS1 Thromboxane synthase 059377 PAF orPCLAF platelet activating factor 166803 cPLA2a or PLA2G1B PhospholipaseA2 170890 THBS1 Thrombospondin 137801 CD40L or CD40LG CD40 ligand 102245SgIII or SCG3 Secretogranin III 104112 COL18A1 Endostatin 182871 TGF-βor TGFB1 transforming growth factor beta 105329 TLN1 Talin1 137076 MOGmyelin oligodendrocyte glycoprotein 232641, 137345, 236561, 137345,204655, 234623, 237834, 234096 GAD2 glutamic acid decarboxylase 2136750, 136750 MAG myelin associated glycoprotein 105695 PMP22peripheral myelin protein 22 109099 TPO thyroid peroxidase 277603,115705 VGKC voltage-gated potassium channel genes N/A PLP or PLP1proteolipid protein 123560 AChR or CHRNA1 acetylcholine receptor 138435AChR or CHRNA10 acetylcholinereceptor 129749 AChR or CHRNA2acetylcholinereceptor 120903 AChR or CHRNA3 acetylcholinereceptor 080644AChR or CHRNA4 acetylcholinereceptor 101204 AChR or CHRNA5acetylcholinereceptor 169684 AChR or CHRNA6 acetylcholinereceptor 147434AChR or CHRNA7 acetylcholinereceptor 175344 AChR or CHRNA9acetylcholinereceptor 174343 AChR or CHRNB1 acetylcholinereceptor 170175AChR or CHRNB1 acetylcholinereceptor 283946 AChR or CHRNB2acetylcholinereceptor 160716 AChR or CHRNB3 acetylcholinereceptor 147432AChR or CHRNB4 acetylcholinereceptor 117971 AChR or CHRNDacetylcholinereceptor 135902 AChR or CHRNE acetylcholinereceptor 108556AChR or CHRNG acetylcholinereceptor 196811 TRIB2 tribbles pseudokinase 2071575 GluR or GRIA1 N-methyl-D-aspartate (NMDA)-type glutamate receptor155511 GluR or GRIA2 N-methyl-D-aspartate (NMDA)-type glutamate receptor120251 GluR or GRIA3 N-methyl-D-aspartate (NMDA)-type glutamate receptor125675 GluR or GRIA4 N-methyl-D-aspartate (NMDA)-type glutamate receptor152578 GluR or GRIK3 N-methyl-D-aspartate (NMDA)-type glutamate receptor163873 GAD2 glutamate decarboxylase 2 136750, 136750 ARMC9 Armadillorepeat containing 9 135931 CYP21A2 Cytochrome P450 Family 21 Subfamily AMember 2 231852, 235134, 198457, 232414, 233151, 206338 CASR calciumsensing receptor 036828 NASP nuclear autoantigenic sperm protein 132780INS insulin 254647 TSHR thyroid stimulating hormone receptor 165409 TPOthyroperoxidase 277603, 115705 ASGR1 asioglycoprotein receptor 1 141505ASGR2 asioglycoprotein receptor 2 161944 CYP2D6 Cytochrome P450 Family 2Subfamily D Member 6 272532, 275211, 100197, 280905, 282966, 283284 LFor LTF lactoferrin 012223 TTG or TGM1 tissue trans-glutaminase 1 285348,092295 TTG or TGM2 tissue trans-glutaminase 2 198959 TTG or TGM3 tissuetrans-glutaminase 3 125780 H/K ATP-ase gastric hydrogen potassium ATPaseF8 Factor XIII 185010 Beta2-GPI or APOH beta2-glycoprotein I 091583ITGB2 B2 integrin 160255 G-CSF or CSF3 granulocyte-colony stimulatingfactor 108342 GP IIb/IIa glycoprotein N/A COLII or COL2A1 collagen II139219 MPO myeloperoxidase 005381 CYO or MYH7 cardiac myosin 092054PRTN3 proteinase 3 277804, 196415 TCHH trichohyalin 159450 GP1 or GTPBP1glycoprotein 1 100226 LM332 laminin-332 N/A COLVII COL7A1 type VIIcollagen 114270 COIL P80 Coilin 121058 DSG1 Desmoglein 1 134760 DSG3Desmoglein III 134757 SOX10 SRY-Box 10 100146 70SNRNP70 or RNU1-1 smallnuclear ribonucleoprotein U1 subunit 206652 SAG S-antigen 130561, 281857α3(IVNC1 collagen) or COL4A3 Collagen alpha-3(IV chain) 169031ANO6/TMEM16F Anoctamin 6/Transmembrane Protein 16F 177119

Symbols and names are used herein, along with ENSEMBL protein IDs, torefer to proteins from humans. Unless otherwise noted, the protein name(if used to refer to the protein herein) and symbol and ENSEMBL protein(ENSP) IDs corresponding to each symbol are shown in Table 2. The uniqueidentifiers for each ENSEMBL entry has been modified to remove the firstfive leading zeros (0) of the identifier after the ENSP label.

TABLE 2 ENSEMBL identifiers for antibody targets Symbol ENSP(s) ACVR2Bor ACVR2B 340361 ACVRL1 455848, 373574, 446724, 447884, 392492, 457394AFP 379138, 226359 ANGPT2 or Ang-2 314897, 486858, 343517, 428023ANGPTL3 360170 AOC3 or AOC3 312326, 464787, 465913, 468632, 468043,477686, 484312 APCS 255040 APP or N/a 284981, 346129, 345463, 350578,387483, 398879, 351796, 396923, 406539, 352760 AXL or AXL 301178,351995, 471497 B4GALNT1 n/a BSG or CD147 473664, 344707, 333769, 458665,343809, 495088, 484849, 478958, 484624, 473393, 473528 C1S or C1s385035, 328173, 354057, 384171, 399892, 406643, 384464, 397921, 442298,484657 C5 or C5 223642 CA9 367608, 482050 CALCA 417833, 420618, 331746,379657, 354286 CASP2 312664, 376656, 340030, 481929 CCL11 or CCL11302234 (eotaxin-1) CCL2 or MCP-1 462156, 225831 CCR2 or CCR2 399285,396736, 383681, 292301 CCR4 or CCR4 332659 CCR5 or CCR5 292303, 404881CD19 or CD19 313419, 456201, 437940 CD2 or CD2 358490, 358489 CD200 orCD200 476114, 475860, 418576, 420298, 312766, 373179, 419816 CD22 orCD22 469980, 470681, 469503, 472664, 472762, 471972, 471399, 469523,470193, 442279, 085219, 441237, 403822, 469984, 470724, 482823, 473221,339349 CD27 or CD27 266557 CD274 or PD-L1 370989, 370985 CD276 or CD276455366, 320084, 441087, 453907, 452649, 452736, 454258, 453330, 452905,453336, 456657, 452669, 454940, 453014, 453842, 320058 CD276 or B7-H3n/a CD28 or CD28 393648, 324890, 363605 CD3 or CD3 n/a CD33 or CD33403331, 410126, 375673, 262262 CD37 or CD37 375732, 470394, 413151,471902, 325708, 441037, 471078, 470260, 470683 CD38 or CD38 427277,226279, 423047 CD3E or CD3 epsilon 354566, 433975 CD4 or CD4 011653,445167, 440720 CD40 or CD40 361359, 361350, 434825, 484074 CD40LG orCD154 359663, 359662 (CD40L) CD44 or CD44 v6 263398, 398632, 435377,389830, 432704, 395953, 392331, 432405, 278386, 404447, 309732, 398099,434465, 279452, 434530, 432718, 433189, 436549, 436623, 436980, 278385,435321, 436451, 434418, 431860, 434920, 492449 CD52 or CD52 363330 CD6or CD6 323280, 443748, 340334, 440055, 410638, 390676, 340628, 443747CD70 or CD70 395294, 245903, 470805 CD74 or CD74 367026, 230685, 430614,429024, 009530, 430654, 429641, 429478 CD79B or CD79B 376544, 006750,245862 CD80 or CD80 264246, 418364, 373165 CD97 or CD97B n/a CEACAM5 orCEA 381600, 221992, 385072, 468997, 473252, 482303, 482157, 480800CFAP221 or PCDP1 295220, 470662, 391760, 409912, 470784, 393222, 472563,470283, 472069, 471092, 413299, 471998, 399793 CFD or CFD 478745,488580, 332139, 468253 CLDN18 or CLDN18.2 340939, 183605, 419732 CLEC6A371505 CLTA4 or CD152 n/a CSF1 or CSF1 434527, 431547, 349854, 327513,433837, 358817, 407317, 358816 CSF1R or CSF1R 422212, 286301, 427545,421174, 445282 CSF2 or CSF2 296871 CSF2RA 370940, 370935, 410667,416437, 370920, 476684, 348058, 436825, 347606, 370911, 440491, 394227CTGF or CTGF 356954 CTLA4 or CTLA-4 497102, 303939, 295854, 497319,417779 CXCL10 305651 CXCR4 or CXCR4 386884, 241393 (CD184) DLL3 or DLL3348810, 471688, 205143 DLL4 or DLL4 497860, 249749 DPP4 or DPP4 353731,402259, 401359, 410264, 486421 EGFL7 or EGFL7 360764, 385639, 473338,360763, 307843 EGFR or EGFR 415559, 342376, 345973, 413843, 275493,413354, 492462, 395243 ENG 362299, 341917, 479015 EPCAM or EpCAM 385476,410675, 263735, 389028 EPHA3 337451, 399926, 419190 ERBB2 or HER2462438, 462808, 404047, 463714, 462024, 463427, 269571, 463719, 464420,464252, 463002, 385185, 446466 ERBB3 or ERBB3 (HER3) 495453, 448636,449138, 267101, 415753, 448671, 448483, 447510, 449713, 408340, 482073,448946, 449129, 448729 F3 334145, 359226 F9, F10 n/a FAP or FAP 441940,417028, 188790, 411391, 400137, 407404, 485844 FCER2 or CD23 264072,471974, 472067, 353178 FCGRT or FCGRT 221466, 472350, 472794, 469968,410798, 469933, 471300, 471118, 472604, 472256, 471232 FGB or 306099,398719, 426757 FGF23 or FGF 23 237837 FGFR2 or FGFR2 351276, 474011,491912, 348559, 358056, 474109, 358055, 404219, 263451, 410294, 353262,358052, 358054, 337665, 352309, 481464, 484892, 490905, 350166, 484154,358057, 309878 FLT1 or VEGFR-1 282397, 437631, 484039, 491097, 442630,484385, 443311, 484832, 437841 FN1 394423, 323534, 338200, 350534,346839, 410422, 415018, 399538, 348285, 416139, 392565, 398907, 352696FOLH1 256999, 349129, 434928, 344131, 431463, 436569, 431577, 431263FOLR1 308137, 377286, 377284, 377281 FOLR2 405638, 298223, 414094,443307, 441547, 438568, 444794, 321957, 440337, 480592 FUT4 or CD15351602 FZD1 287934 GCGR or GCGR 383558, 460976, 458930 GPC3 359854,486325, 385307, 377836 GPNMB or GPNMB 258733, 371420, 386476, 497362GUCY2C or GUCY2C 261170 HGF or HGF 494899, 222390, 391238, 389854,408270, 413829, 494355, 346164, 496217, 396307, 388592 HLA-DRA or HLA-372746, 372745, 404533, 392789, 410443, 411610, 479287, 405295, 398838,378786, DR 364121, 372608, 403385, 402951, 412562 HSP90AA1, 216281,335153, 451400, 450712, 452241, 489370 HSP90AA2, HSP90AB1, HSOP90B1,TRAP1 or Hsp90 ICAM1 or 1CAM-1 264832, 413124, 465680 ICOS or CD278, aka319476, 415951 ICOS ICOSLG or ICOSL 494882, 339477, 384432, 383230,383228 IFNA1 or IFN-α 276927 IFNAR1, IFNAR2 270139, 400161 IFNG 229135IGF1R or CD221 497069, 496919, 268035, 453007, 453630, 454115, 456950IGHE or IGHE 492979, 374983, 481089 IL12A or IL-12 303231, 420184,419046 IL13 or IL-13 304915, 479835 IL17A or IL17A n/a IL17F or IL17F337432 IL1A or IL1A 263339 IL1B or IL-1β 263341, 407219, 409680, 400854IL2 or IL2 226730 IL20 or IL 20 356065, 356063, 375796 IL22 or IL-22442424, 329384 IL23A or IL23 228534 IL2RA or CD25 369293, 369287,256876, 402024 IL31RA 380048, 380046, 415900, 351935, 427533, 347047,297015, 479432 IL3RA or IL 3 receptor 327890, 414867, 370878 IL4 or IL4231449, 325190, 480581 IL5 or IL-5 231454, 409825 IL5RA or CD125 412209,390753, 256452, 373358, 309196, 400400, 392059, 398117, 391274, 388858IL6 or IL6 385675, 405150, 385718, 385043, 384928, 385227, 258743 IL6Ror IL-6R 357470, 340589, 423184, 423668, 423036, 477739 IL9 or IL9274520 ITGA2 or ITGA2 296585, 426489, 422095, 424397, 424642, 422145(CD49b) ITGA2B or CD41 498119, 467269, 262407 ITGA5 293379, 450267,405865, 447347 ITGAL or LFA-1 349252, 456521, 350886, 457785, 454908,454342, 456888, 409377, 461006, 458739 (CD11a) ITGAV or CD51 261023,364042, 404291, 389442 ITGB2 or ITGB2 380950, 380955, 380952, 347279,380948, 427732, 317697, 428503, 428979, 428413, (CD18) 428125, 428434,430901, 428870, 380944, 429683, 303242 ITGB3 452786, 461626, 465586ITGB7 267082, 456446, 408741, 455374, 437375, 450366, 456305, 446703 KDRor VEGFR2 495159, 263923 KIR2DL1, KIR2DL2 480247, 484701, 477690,479363, 484700, 482506, 478633, 484559, 481123, 478054, or KIR2D 478567,480989, 479574, 482449, 336769, 291633, 478202, 484361, 479644, 483525,478232, 481722, 478604, 484871, 478895, 482501, 478263, 482120, 479941,484582, 482456, 481187, 492250, 492549, 492477, 491211, 491668, 492815,491499, 492598, 492436, 492695, 491930, 491721, 491975, 492286, 491348,492859 KLRC1 or NKG2A 441432, 352064, 385304, 256965, 442545, 438038LAG3 or LAG3 413825, 203629 LINGO1 or LINGO-1 347451, 453853, 453780,455605, 454465, 454687, 454051, 454245, 454577, 456516, 457101 LOXL2 orLOXL2 373783, 473322, 427907, 427826, 427883, 429778, 430519, 428497,428933 LRRC15 or LRRC15 306276, 413707 LTA or LTA 403495, 413450,372791, 372793, 383131, 372991, 395976, 416509, 395895, 407133, 416337,387924, 412555, 402413 LYPD3 or LYPD3 244333 MADCAM1 475575, 304247,215637, 372130, 480908, 484317, 480104, 483663, 484153 MAG 470772,376048, 355234, 473125, 473245, 440695 MAPT 487613, 488245, 482244,488081, 487819, 488373, 488046, 487837, 488101, 484491, 484321, 478602,483396, 480217, 486039, 487570, 485913, 477703, 481769, 487403, 483784,485831, 479142, 460048, 334886, 408975, 413056, 410838, 458742, 460965,389250, 262410, 303214, 340820, 443028, 340438 MASP2 or MASP-2 383690,383691 MCAM or MCAM 264036 MET 413857, 380860, 317272, 398776, 398140,410980 MIF or MIF 482779, 215754 MMP9 361405 MS4A1 or CD20 433179,432219, 433519, 433277, 432270, 437002, 314620, 374589 MSLN 456008,442965, 456702, 372313, 456132, 458003, 454295, 457847 MST1R or MST1R296474, 407926, 341325, 393294, 414792, 482642, 481084, 482827 (aka RON)MSTN or GDF-8 260950 MUC1 481231, 479471, 478068, 357377, 389098,482688, 357378, 357374, 357381, 339690, 342814, 483128, 484006, 484730,357383, 357375, 338983, 483482, 343482, 483473, 484824, 483581, 388172,482988, 480335, 480333 MUC16 or CA-125 381008, 472883, 470885, 472781MUC5AC or 5AC 485659, 490794 MYH7 347507 NCAM1 or CD56 480132, 384055,481083, 479353, 318472, 482852, 484943, 482205, 480774, 484481, 479687,475074, 486406, 480269, 478072, 486241, 477835, 480797, 477808, 479241NECTIN4 356991 NGF or HNGF 358525 NOTCH1 or Notch 1 277541 NRP1 or NRP1364009, 265371, 390447, 416147, 476896, 393071, 363954, 363955, 363956,408911, 390567, 379317, 364001, 363949 NT5E 358660, 257770, 414674,387630, 358665 PCDHAC1 or PCDC1 386356, 253807 PCSK9 or PCSK9 303208PDCD1 or PD-1 480684, 486779, 487175, 335062, 390296, 340808 PDGFRA orPDGF-Rα 257290, 425648, 425626, 424218, 425902, 426472, 425232 PDGFRB261799, 430026, 429218, 430715 PTDSS1 430548, 337331, 430928 PTK7 orPTK7 418386, 420186, 418462, 419096, 418545, 420765, 420165, 418754,230418, 325992, 326029, 325462, 419037, 420322, 230419, 417607 PTPRC orCD45 356349, 494132, 306782, 411355, 433536, 494327, 356337, 356334,405494, 469141, 393360, 458846, 461347, 458322, 458662, 461074, 458191,458418, 482203, 461712, 483380 RGMA or RGMA 330005, 440025, 451505,452126, 452170, 456290, 451709, 452350, 404442, 442498 RHD or RHD331871, 498055, 413849, 396420, 350150, 456966, 339577, 399640, 478087RHD, RHCE 294413, 345084, 311185, 431741, 344485, 334570, 435401,415417, 331871, 498055, 413849, 396420, 350150, 456966, 339577, 399640,478087 ROR1 or ROR1 360121, 360120, 441637 RSPO3 349131, 357300 RTN4 orRTN4 378107, 384471, 349944, 337838, 322147, 350365, 378109, 385650,489133, 397808 411628, 384825 S1PR1 498194, 305416, 497175, 498038,497478 SAA1, SAA2 or 348918, 436866, 497498, 384906 SDC1 or SDC1 254351,384613, 400773, 390201, 370542 SELL or CD62L 236147 SELP 356769, 356762,399368, 356760, 263686, 391694 SLAMF7 or CD319 357022, 357021, 352281,473590, 403294, 416592, 409965, 405605, 483774 SLC39A6 or LIV-1 269187,465915, 401139, 467724 SLITRK6 or SLITRK6 495507, 383143, 496428 SNCA orNACP 338345, 343683, 378437, 378442, 426955, 422238, 421485, 479604,426034, 423445, 396241, 484044, 378440 SOST or SOST 301691 ST8SIA1379353, 261197, 441707, 444999, 440292, 384467, 370832 STEAP1 or STEAP1297205, 394402 TACSTD2 or TROP-2 360269 TFPI or TFPI 376172, 233156,409177, 386344, 342306, 388159, 408170, 394185, 400179, 402954 TGFB1 orTGF-β 472767, 221930 TGFB2 or TGF beta 2 355897, 355896 TIGIT or TIGIT418917, 420552, 419085, 419706, 373167 TNC 265131, 339553, 411406,443478, 442242, 445380, 489385, 443469, 438152 TNF or TNF-α 398698,365290, 389492, 389490, 392858, 389265, 372988, 410668 TNFRSF10A or221132, 428884, 480778 TRAIL-R1 TNFRSF10B or 276431, 317859, 427999TRAIL-R2 TNFRSF12A 458898, 326737, 343894, 458305, 460610, 461756TNFRSF13C or BAFF-R 291232 TNFRSF17 or BCMA 053243, 379753, 454314TNFRSF4 or OX-40 368538 TNFRSF8 or CD30 263932, 421938, 398337, 390650(TNFRSF8) TNFRSF9 or 4-1BB 366729, 465272, 464978, 478699 (CD137)TNFSF11 or RANKL 351347, 381775, 384042, 444913, 239849 TNFSF13B or BAFF365048, 389540, 445334 TPBG or 5T4 358765, 440049, 441219, 489447,489143, 489140 TRAP or TRAP n/a TSLP or TSLP 399099, 339804, 427827TYRP1 or TYRP1 419006, 373570, 370528 VEGFA or VEGF-A 361137, 317598,388663, 389864, 361125, 421561, 388465, 361134, 361148, 430594, 428321,430479, 429643, 409911, 430829, 429008, 430002, 230480, 429592, 478570,483241, 484284, 492199, 478034, 492413, 492800 VIM 446007, 489830,490509, 224237, 435613, 431702 VSIR or VSIR 378409 VWF or VWF 261405,461331, 459134 TAG-72 n/a

CD3 or CD3 is also known as Cluster of differentiation 2 (multiplesubunits). FCER2 or CD23 is also known as (IgE receptor. NT5E is alsoknown as 5′-nucleotidase. F9, F10 is also known as activated F9, F10.ACVRL1 is also known as activin receptor-like kinase 1. AFP is alsoknown as alpha-fetoprotein. ANGPTL3 is also known as angiopoietin 3. BSGor CD147 is also known as basigin. APP or N/a is also known asbeta-amyloid. CALCA is also known as calcitonin gene-related peptide.CA9 is also known as carbonic anhydrase 9 (CA-IX). MYH7 is also known ascardiac myosin. MET is also known as c-Met. F3 is also known ascoagulation factor III. CLEC6A is also known as dendriticcell-associated lectin 2. EGFR or EGFR is also known as elongatinggrowth factor receptor. ENG is also known as endoglin. EPHA3 is alsoknown as ephrin receptor A3. FGB or is also known as fibrin II, betachain. FN1 is also known as fibronectin extra domain-B. FOLH1 is alsoknown as folate hydrolase. FOLR2 is also known as folate receptor 2.FOLR1 is also known as folate receptor alpha. FZD1 is also known asFrizzled receptor. B4GALNT1 is also known as GD2 ganglioside. ST8SIA1 isalso known as GD3 ganglioside. MMP9 is also known as gelatinase B. TYRP1or TYRP1 is also known as glycoprotein 75. GPC3 is also known asglypican 3. CSF2RA is also known as GMCSF receptor α-chain. IGF1R orCD221 is also known as IGF-1 receptor. IL31RA is also known as IL31RA.ITGA2B or CD41 is also known as integrin alpha-IIb. ITGA5 is also knownas integrin α5. ITGB3 is also known as integrin αIIbβ3. ITGB7 is alsoknown as integrin β7. IFNG is also known as interferon gamma. IFNAR1,IFNAR2 is also known as interferon α/β receptor. CXCL10 is also known asinterferon gamma-induced protein. IL12A or IL-12 is also known asinterleukin 12. IL13 or IL-13 is also known as interleukin 13. IL17A orIL17A is also known as interleukin 17 alpha. IL17F or IL17F is alsoknown as interleukin 17F. IL2 or IL2 is also known as interleukin 2.IL22 or IL-22 is also known as interleukin 22. IL23A or IL23 is alsoknown as interleukin 23. IL6 or IL6 is also known as interleukin 6. SELLor CD62L is also known as L-selectin. MSLN is also known as mesothelin.MUC1 is also known as mucin CanAg. MADCAM1 is also known as mucosaladdressin cell adhesion molecule. MAG is also known as myelin-associatedglycoprotein. NECTIN4 is also known as nectin-4. CASP2 is also known asneural apoptosis-regulated proteinase 2. PTDSS1 is also known asphosphatidylserine. PDGFRB is also known as platelet-derived growthfactor receptor beta. RHD, RHCE is also known as Rhesus factor. RSPO3 isalso known as root plate-specific spondin 3. SELP is also known asselectin P. SAA1 or SAA2 is also known as serum amyloid A protein. APCSis also known as serum amyloid P component. SIPR1 is also known assphingosine-1-phosphate. MAPT is also known as tau protein. TNC is alsoknown as tenascin C. TNFRSF12A is also known as TWEAK receptor. VIM isalso known as vimentin. VWF is also known as von Willebrand factor.IL2RA or CD25 is also known as α chain of IL-2receptor.

III. Compositions of the Invention

Various embodiments of the inventions described herein provideengineered megakaryocyte progenitors to encoding a chimeric plateletreceptor (CPR). The receptor may bind a specific antigen or targetpresent on a tumor or specific location in the body, for example theantigen to which the CPR binds may be an endogenous antigen. In someembodiments the target is not collagen. Alternatively, plateletreceptors may be deleted to prevent cargo or toxin release in responseto normal platelet activation signals. The multiple edits required togenerate these progenitor cells are possible because the progenitorcells never enter the patient's body where there are concerns ofcontinuous culture or genome instability. Only enucleated platelets areinjected into the patient.

A. Engineering Megakaryocytes

In some embodiments, the engineered platelets described herein originatefrom genetically modified megakaryocytes. The genome of thesemegakaryocytes may include a knock-out of at least one, two, three,four, five, six, seven, eight, nine, or at least ten genes encoding anendogenous receptor, mediator protein, and/or signaling transductionprotein. It will be clear that in some instances it may not be necessaryto knock out or delete the entire gene. For example GP1b knockoutresults in abnormal platelets, however one can delete only theextracellular domain of the receptor (removing its ability to function)while retaining the intracellular domain, resulting in typical plateletsthat lack the ability to bind to von Willebrand factor the GP1b target).Accordingly in some embodiments, the disruptions, deletions or knockoutsdescribed herein are full disruptions, deletions or knockouts of theentire gene. In other embodiments, the disruptions, deletions andknockouts are disruptions deletions and functional knockouts i.e.disruption of the function of the protein, and in some embodiments thedeletion is a deletion of the extracellular domain of the proteins.

Examples of genes that may be deleted from the megakaryocyte genome areshown in Table 3.

TABLE 3 Potential genes for deletion Gene Mediator or receptor? GeneMediator or receptor? Rab27a Mediator Factor XIII mediator HPS genesMediator PF4 mediator integrin AIIbB3 receptor NAP2 (Nucleosome mediatorAssembly Protein 2) GP1b-IX-V receptor Prothrombin mediator Par1receptor High Molecular Weight mediator Kininogens Par4 receptorPlasminogen activator mediator inhibitor 1 P2Y1 receptor a2-antiplasminmediator P2Y12 receptor plasminogen mediator IP receptor P-Selectinmediator/receptor TP receptor CXCL4 mediator TLR (many) receptor CXCL7mediator GPV1 receptor FGF mediator a2B1 (type 1 collagen receptor)receptor EGF mediator GPIIbIIIA HGF CLEC-2 receptor IGF MyD88 (MyeloidDifferentiation signal transduction Angipoetin Primary Response 88)Galphaq signal transduction Thromboxane synthase mediator LIMK1 mediatorPAF Mediator vWF mediator cPLA2a mediator Fibrinogen mediatorThromospondin PDGF mediator CD40L VEGF mediator SgIII (SecretograninIII) Factor V mediator Endostatin Factor VIII mediator TGF-β(transforming growth factor beta) Factor XI mediator Talin1 signaltransduction ANO6/TMEM16F mediator Kindlins signal transductionThe skilled person will appreciate that there are several pathways whichshould be disrupted to allow the production of a platelet with reducedthrombogenic potential. In some embodiments any one or more of thefollowing three pathways are disrupted: recognition of primary stimuliof thrombus formation; recognition of secondary stimuli of thrombusformation; and release of secondary mediators of thrombus formation.Recognition of primary stimuli of thrombus involves the plateletsrecognizing factors associated with exposed tissue that becomes exposedupon wounding, for example, recognizing the subendothelium. In typicallycircumstances, platelets are not exposed to subendothelium. Exposure ofthe subendothelium allows platelets to recognize ligands such ascollagen, von Willebrand factor, fibronectin, thrombospondin viareceptors on the platelet surface, such as GPIb/V/IX and GPVI (GP6),ITGA2B, integrins s α_(IIB)β₃, α₂β₁, α₅β₁ and α₆β₁. Accordingly, in someembodiments the genes encoding a protein involved in recognition ofprimary stimuli of thrombus formation include GPIb/V/IX and GPVI (GP6),ITGA2B, CLEC2, integrins s α_(IIb)β₃, α₂β₁, α₅β₁ and α₆β₁.Once platelets have made contact with the exposed endothelium, forexample via the interactions discussed above, the platelets releasesecondary messengers such as ADP, thrombin and TxA₂ which are detectedby other platelets and which cause platelet aggregation at the woundsite. In some embodiments, it is preferred if the ability of theplatelets to recognize the secondary messengers is disrupted. It is notdesirable if a platelet of the invention is targeted to wound site forexample, rather than the intended target. Accordingly, in preferredembodiments the ability of the platelets to recognize the secondarymessengers is disrupted. Receptors that are involved in this functioninclude Par1, Par4, P2Y12, GPIb/V/IX, the Thromboxane receptor (TBXA2R),P2Y1, P2X1 and integrin α_(IIb)β₃.As mentioned above, once platelets have recognized the exposed tissue,they release secondary messengers to recruit other platelets to thesite. Once a platelet of the invention has bound to a target, forexample to a tumour antigen, it is not desirable for the platelet of theinvention to then recruit other platelets to a target site and form athrombus, for example a thrombus at a tumour site.Accordingly, in preferred embodiments, the pathway by which theactivated platelet releases the secondary messengers is disrupted. Thepathway can include those proteins that are involved in the productionand/or storage and/or release of the secondary mediators. Genes involvedin this pathway include Cox1, HPS and thromboxane-A synthase (TBXAS1).The skilled person will appreciate that a single gene can be involved intwo or three of the above functions.In some embodiments it is preferred if the megakaryocyte or progenitorthereof comprises a disruption or deletion of (e.g. a knockout of) atleast one, two, three, four, five, six, seven, eight, nine, or at leastten genes encoding a protein involved in recognition of primary stimuliof thrombus formation; a protein involved in recognition of secondarymediators of thrombus formation; and/or a protein involved in therelease of secondary mediators of thrombus formation.It will be clear to the skilled person that by a protein involved inrecognition of primary stimuli of thrombus we include the meaning of anyprotein that is involved in this process, for example includes theprotein that is directly involved in contact with or recognition ofprimary stimuli of thrombus, and also genes that for example lead to theexpression of those proteins that are directly involved in contact withor recognition of the primary stimuli of thrombus. The skilled personwill understand which proteins are considered to be involved inrecognition of primary stimuli. The key feature is that disruption ofthe proteins are that their disruption leads to a defect in therecognition of primary stimuli of thrombus. However, in some embodimentsa protein involved in recognition of primary stimuli of thrombusincludes only those proteins that directly make contact with the primarystimuli of thrombus.By a protein involved in recognition of secondary mediators of thrombusformation we include those proteins that are directly involved in thecontact with or recognition of secondar mediators of thrombus formation,as well as proteins that are indirectly involved in those processes, forexample those proteins that are involved in the production of theproteins that are directly involved in the contact with or recognitionof secondar mediators of thrombus formation. The skilled person willunderstand what is mean by proteins involved in recognition of secondarymediators of thrombus formation. The key feature of the proteins arethat their disruption leads to a defect in the recognition of secondarymediators of thrombus formation. However, in some embodiments a proteininvolved in recognition of secondary mediators of thrombus formationincludes only those proteins that make direct contact with the secondarymediators of thrombus formation.By a protein involved in the release of secondary mediators of thrombusformation we include those proteins that are involved in the productionand/or storage and/or release of the secondary mediators. The keyfeature of the proteins are that their disruption leads to a defect inthe ultimate release of the secondary mediators. The defect may be inthe production of the secondary mediators, the storage of the secondarymediators, and/or the actual release process.In some embodiments the megakaryocyte or progenitor thereof comprises adisruption or deletion of at least:

one gene that encodes a protein involved in recognition of primarystimuli of thrombus formation;

one gene that encodes a protein involved in recognition of secondarymediators of thrombus formation; and

one gene that encodes a protein involved in the release of secondarymediators of thrombus formation;

In some embodiments the megakaryocyte or progenitor thereof comprises adisruption or deletion of at least:

two genes that encode a protein involved in recognition of primarystimuli of thrombus formation;

two genes that encode a protein involved in recognition of secondarymediators of thrombus formation; and

two genes that encode a protein involved in the release of secondarymediators of thrombus formation:

In some embodiments the megakaryocyte or progenitor thereof comprises adisruption or deletion of at least:

three genes that encode a protein involved in recognition of primarystimuli of thrombus formation;

three genes that encode a protein involved in recognition of secondarymediators of thrombus formation; and

three genes that encode a protein involved in the release of secondarymediators of thrombus formation.

-   -   Genes that are considered to encode a protein involved in        recognition of primary stimuli of thrombus formation include        GPIb/V/IX and GPVI (GP6), ITGA2B, CLEC2, integrins s α_(IIb)β₃,        α₂β₁, α₅β₁ and α₆β₁, or optionally include GPVI and ITGA2B.    -   Genes that are considered to encode a protein involved in        recognition of secondary stimuli of thrombus formation include        Par1, Par4, P2Y12, GPIb/V/IX, the Thromboxane receptor (TBXA2R),        P2Y1, P2X1 and integrin α_(IIb)β₃ or optionally include Par1,        Par4 and P2Y12.    -   Genes that are considered to a protein involved in release of        secondary mediators of thrombus formation include Cox1, HPS and        thromboxane-A synthase (TBXAS1), or optionally include Cox1 and        HPS.        In some embodiments    -   the at least one, two or three genes that encode a protein        involved in recognition of primary stimuli of thrombus formation        are selected from the group consisting of: GPIb/V/IX and GPVI        (GP6), ITGA2B, CLEC2, integrins s α_(IIb)β₃, α₂β₁, α₅β₁ and        α₆β₁, or from the group consisting of GPVI and ITGA2B;    -   the at least one, two or three that encode a protein involved in        recognition of secondary mediators of thrombus formation are        selected from the group consisting of Par1, Par4, P2Y12,        GPIb/V/IX, the Thromboxane receptor (TBXA2R), P2Y1, P2X1 and        integrin α_(IIb)β3 or from the group consisting of Par1, Par4        and P2Y12; and/or    -   the at least one, two or three genes that encode a protein        involved in the release of secondary mediators of thrombus        formation are selected from the group consisting of Cox1, HPS        and thromboxane-A synthase (TBXAS1) or from the group consisting        of Cox1 and HPS.        In a preferred embodiment, the genetically modified        megakaryocyte or progenitor thereof has a disruption or deletion        in each of the following genes:

GPVI, ITGA2B, Par1, Par4, P2Y12, Cox1 and HPS.

For example the genetically modified megakaryocyte or progenitor thereofmay comprise a knockout of each of GPVI, ITGA2B, Par1, Par4, P2Y12, Cox1and HPS.

In some embodiments, expression of the genes in Table 3 may be alteredor “knocked-out” using a CRISPR/Cas system, zinc finger nucleases,transcription activator-like effector nucleases (TALENs), a RNAinterference construct (RNAi) (e.g., small interfering RNA (siRNA) ormicroRNA (miRNA)), or a short hairpin RNA (shRNA).

The effects of knock-out of a gene in a megakaryocyte on the resultingengineered platelet may be varied. For example, RAB27a (RAS oncogene)and HPS (haptoglobin) genes function in dense granule loading andformation, respectively. Knock-out or deletion of Rab27a may result inengineered platelets with no dense granule mediators but with otherwisenormal platelet biology. Knock-out or deletion of HPS genes may resultin engineered platelets containing no dense granules. Knock-out ordeletion of AIIbB3 or GP1b-IX-V may result in failure of the plateletsto aggregate with each other by decreasing interaction between theplatelet and von Willebrand factors (vWF) after activation. Further,AIIbB3 is also involved in inside-out signaling to increase the affinityof the integrin for fibrinogen (See, Durrant, Blood. 2017 Oct. 5;130(14): 1607-1619). Knock-out or deletion of IP (PGI2R or prostaglandin12 receptor) may result in negative regulation of prostaglandin.Knock-out or deletion of TP (TxA2R or Thromboxane A2 Receptor) mayresult in reduction of recruitment of additional platelets on activationto stimulate clotting.

GPVI (ITAM receptor) has been observed to still be stimulated inG-protein alpha-q (Galphaq) knockout mice. Conversely, ITAM agonists,such as collagen, induce release of G-protein-coupled receptors (GPCRagonists), such as ADP and thromboxane A2 receptor (TXA2), thusindirectly activating phospholipase C (PLC) through the Gq pathway.Further, Galphaq is active for proper function for thrombin, ADP,5-hydroxytryptamine (5HT), PAF, and thromboxane A (TXA).

Knock-out or deletion of P-selectin, thromboxane synthase, and plateletactivating factor (PAF) may result in failure of platelet aggregationonce activated. Knock-out or deletion of LIM Domain Kinase 1 (LIMK1)will likely reduce TxA2 synthesis. CXCL4 (C-X-C motif chemokine ligand4) and CXCL7 (C-X-C motif chemokine ligand 7) are chemokines; therefore,knock-out or deletion of the gene would likely interfere in at least onesignaling pathway. Talin1 and kindlins function in signal transductionto allow integrins to enter a sensitive state.

Knock-out or deletion of ANO6/TMEM16F disrupts the platelets ability toexpose phosphatidylserine on platelet activation. Phosphatidlyserine isa membrane lipid which is usually kept on the cytoplasmic face of theplatelet. On platelet activation, calcium influx triggersphosphatidylserine exposure on the outside of the platelet viaANO6/TMEM16F, where it acts to catalyse the production of activethrombin in combination with clotting factors. Thus, knockout of TMEM16Fprevents phosphatidylserine exposure and thus would decrease plateletthrombogenicity. This is exemplified by Scott's syndrome patients, whofeature ANO6 mutations and clinically have increased risk of bleeding.

B. Chimeric Platelet Receptors (CPR)

In some embodiments, the engineered platelets described herein mayinclude alterations to the endogenous platelet receptors. Alterationsinclude, but are not limited to, deletions or additions or entirereceptors or domains of these receptors, or combinations with domainsfrom non-endogenous receptors to result in differences in the behaviorof an engineered platelet compared to a platelet without the alteration.To stimulate activation of platelets, domains from an immunoreceptortyrosine-based activation motif (ITAM) receptor may be used in achimeric platelet receptor. To inhibit activation of plateletsimmunoreceptor tyrosine-based inhibition motif (ITIM). It will be clearto the skilled person that domains from an ITAM receptor that is nottypically expressed in platelets will still function in the invention,since the ITAM domains are still capable of activating the samedownstream signaling components as ITAM receptors are endogenously foundin platelets.

1. Endogenous Platelet Receptors

In some embodiments, platelets may be redirected to degranulate by anantigen, rather than collagen. ITIM containing receptors inhibitplatelet activation to directly counteract ITAM receptor activation.CEACAM-1, PECAM-1, and G6b-B are ITIM containing receptors. G6b-Bclustering by antibody inhibits platelet activation through GPVI andCLEC-2 as shown in Mori et al. “G6b-B inhibits constitutive andagonist-induced signaling by glycoprotein VI and CLEC-2”. JBC, 2008,which is hereby incorporated by reference in its entirety. Adding achimeric “off” receptor may be used to improve specificity of thesynthetic platelets described herein. An engineered chimericimmunoreceptor tyrosine-based inhibition motif (ITIM) receptor wouldallow logic gate construction.

Alternatively, ITAM receptors mediate platelet activation and stimulatean immune response. Glycoprotein VI (GPVI) binds to collagen and is acentral mediator of platelet activation. It features extracellular IgGlike domains, and the internal tyrosine kinase signaling pathway istriggered by receptor clustering through the Fc receptor (FcR) gammachain. In certain embodiments, the intracellular domain is retained andthe extracellular domain is swapped to target an antigen. For example,in some embodiments, the chimeric platelet receptor comprises anintracellular domain that stimulates platelets, but does not comprisethe corresponding extracellular domain. For example in some embodimentsthe extracellular targeting domain of the receptor is heterologous tothe intracellular domain of the receptor. By heterologous extracellulartargeting domain we mean that the extracellular domain is not the usualextracellular domain associated with the intracellular domain. Forinstance, in embodiments where the intracellular domain comprises theintracellular domain of Glycoprotein VI (GPVI), the extra cellulardomain is not the extracellular domain of Glycoprotein VI (GPVI), thedomains are heterologous to one another.

Alternatively, C-type lectinlike receptor 2 (CLEC-2) or Fc Fragment ofIgG Receptor IIa (FCgR2A) may be altered in a similar way. In otherembodiments, where the intracellular domain comprises the intracellulardomain of C-type lectinlike receptor 2 (CLEC-2), the extracellulartargeting domain is not the extracellular domain of CLEC-2; and in someembodiments where the intracellular domain comprises Fc Fragment of IgGReceptor IIa (FCgR2A), the extracellular targeting domain does notcomprise the extracellular domain of FCgR2A. It is clear that theextracellular targeting domain may be a domain that is native to thesubject, but is not native to the intracellular domain.It will be clear then that in some embodiments the CPR is not anaturally occurring protein.

CLEC-2 binds to podoplanin (associated with tumors) and triggersplatelet activation in response to snake venom rhodocytin and elicitsaggregation of platelets through activation of Src and Syk non-receptortyrosine kinases in the internal tyrosine kinase signaling pathwaytriggered by receptor clustering through signaling proteins lymphocytecytosolic protein 2 or SH2 domain containing leukocyte protein of 76 kDa(SLP-76) and 1-phosphatidylinositol-4,5-bisphosphate phosphodiesterasegamma-2 (PLCγ2) (See, Fu et al. Blood, 127(13):1629-30, 2016, which ishereby incorporated by reference in its entirety).

FCgr2A binds to Fc region of antibodies and triggers platelet activationin response to opsonized bacteria through signaling protein Syknon-receptor tyrosine kinase. The internal tyrosine kinase signalingpathway is triggered by receptor clustering.

In some embodiments, additional ITAM receptors may be inserted into thegenome of a megakaryocyte to enhance T cell signaling and stimulate animmune response. T cell receptors (TCRs) recognize antigens bound in themajor histocompatibility complex (MHC) (See, James et al. Sci. Signal.11, eaan1088 (2018), which is hereby incorporated by reference in itsentirety). ITAMs on the TCRs convert the action of binding andrecognition into an intracellular signal (Ibid). Inserting additionalITAMs into chimeric TCRs was observed to scale linearly with the numberof ITAM receptors and decreasing or knocking-out the number of ITAMreceptors was observed to inhibit T cell development by impairingthymocyte lineage commitment (Ibid).

In some embodiments, a CPR may include one or more domains or portionsthereof from one or more immunoreceptor tyrosine-based activation motif(ITAM) receptors. Non-limiting examples of ITAM receptors includeglycoprotein VI platelet (GPVIA), high affinity immunoglobulin epsilonreceptor subunit gamma (FCERG), C-Type lectin domain family 1 (CLEC1),and Fc fragment of IgG receptor 11 (FCGR2).

In one embodiment, domains of ITAM receptors FCERG (SEQ ID NO: 1), CLEC1(SEQ ID NO: 6), FCGR2 (SEQ ID NO: 10), and/or GPVIA (SEQ ID NO: 15)shown in Table 4 may be combined for expression in a megakaryocyteresulting in a CPR in the engineered platelet.

In one embodiment, domains of ITAM receptors may be combined with T cellreceptor domains to form chimeric ITAM receptors which are also referredto as chimeric platelet receptors. These chimeric receptors may becombined for expression in a megakaryocyte resulting in a CPR in theengineered platelet. Non-limiting examples of chimeric ITAM receptorsfor FCERG (SEQ ID NO: 20), CLEC1 (SEQ ID NO: 21), FCGR2 (SEQ ID NO: 22)and GPVIA (SEQ ID NO: 23) are shown in Table 4.

TABLE 4 ITAM receptors and ITAM chimeric receptors Sequence SymbolRegions Identifier FCERG Signal Peptide (SEQ ID NO: 2); EC Domain (SEQID NO: 3); TM 1 Domain (SEQ ID NO: 4); CytoDomain (SEQ ID NO: 5); stopCLEC1 CytoDomain (SEQ ID NO: 7); TM Domain (SEQ ID NO: 8); EC 6 Domain(SEQ ID NO: 9); stop FCGR2 Signal Peptide (SEQ ID NO: 11); EC Domain(SEQ ID NO: 12); TM 10 Domain (SEQ ID NO: 13); CytoDomain (SEQ ID NO:14); stop GPVIA Signal Peptide (SEQ ID NO: 16); EC Domain (SEQ ID NO:17); TM 15 Domain (SEQ ID NO: 18); CytoDomain (SEQ ID NO: 19); stopFCERG Signal Peptide (SEQ ID NO: 2); Fv1HChain domain (SEQ ID NO: 50);20 (chimeric) Whitlow_Linker (SEQ ID NO: 49); Fv1_Lchain (SEQ ID NO:48); Modified_Hinge_IGg4 (SEQ ID NO: 51); EC Domain (SEQ ID NO: 3); TMDomain (SEQ ID NO: 4); CytoDomain (SEQ ID NO: 5); stop CLEC1 CytoDomain(SEQ ID NO: 7); TM Domain (SEQ ID NO: 8); EC 21 (chimeric) Domain (SEQID NO: 53); Modified_Hing_IGg4 (SEQ ID NO: 51); Fv1HChain (SEQ ID NO:50); Whitlow_Linker (SEQ ID NO: 49); Fv1_Lchain (SEQ ID NO: 48); stopFCGR2 Signal Peptide (SEQ ID NO: 11); Fv1HChain (SEQ ID NO: 50); 22(chimeric) Whitlow_Linker (SEQ ID NO: 49); Fv1_Lchain (SEQ ID NO: 48);Modified_hinge_IGg4 (SEQ ID NO: 51); EC Domain (SEQ ID NO: 54); TMDomain (SEQ ID NO: 13); CytoDomain (SEQ ID NO: 14); stop GPVIA SignalPeptide (SEQ ID NO: 16); Fv1HChain (SEQ ID NO: 50); 23 (chimeric)Whitlow_linker (SEQ ID NO: 49); Fv1_Lchain (SEQ ID NO: 48);Modified_hinge_IGg4 (SEQ ID NO: 51); EC Domain (SEQ ID NO: 55); TMDomain (SEQ ID NO: 18); CytoDomain (SEQ ID NO: 19); stop

SEQ ID NO: 1 is an embodiment of a complete FCERG receptor. SEQ ID NO: 2is an embodiment of the signal peptide of FCERG. SEQ ID NO: 3 is anembodiment of the extracellular domain of FCERG. SEQ ID NO: 4 is anembodiment of the transmembrane domain of FCERG. SEQ ID NO: 5 is anembodiment of the cytoplasmic domain of FCERG. SEQ II) NO: 6 is anembodiment of the ITAM receptor of CLEC1. SEQ ID NO: 7 is an embodimentof the cytoplasmic domain of CLEC 1. SEQ ID NO: 8 is an embodiment ofthe transmembrane domain of CLEC 1. SEQ ID NO: 9 is an embodiment of theextracellular domain of CLEC1. SEQ ID NO: 10 is an embodiment of theITAM receptor of FCGR2. SEQ ID NO: 11 is an embodiment of the signalpeptide of FCGR2. SEQ ID NO: 12 is an embodiment of the extracellulardomain of FCGR2. SEQ ID NO: 13 is an embodiment of the transmembranedomain of FCGR2. SEQ ID NO: 14 is an embodiment of the cytoplasmicdomain of FCGR2. SEQ ID NO: 15 is an embodiment of the ITAM receptor ofGPVIA. SEQ ID NO: 16 is an embodiment of the signal peptide of GPVIA.SEQ ID NO: 17 is an embodiment of the extracellular domain of GPVIA. SEQID NO: 18 is an embodiment of the transmembrane domain of GPVIA. SEQ IDNO: 19 is an embodiment of the cytoplasmic domain of GPVIA.

SEQ ID NO: 20 is an embodiment of a chimeric ITAM receptor based onFCERG. SEQ ID NO: 21 is an embodiment of a chimeric ITAM receptor basedon CLEC1. SEQ ID NO: 22 is an embodiment of a chimeric ITAM receptorbased on FCGR2. SEQ ID NO: 23 is an embodiment of a chimeric ITAMreceptor based on GPVIA.

In some embodiments, a CPR may include one or more domains or portionsthereof from one or more immunoreceptor tyrosine-based inhibition motif(ITIM) receptors. Non-limiting examples of ITIM receptors includeplatelet and endothelial cell adhesion molecule 1 (PECAM1), triggeringreceptor expressed on myeloid cells like 1 (TLT1), leukocyteimmunoglobulin like receptor B2 (LILRB2), carcinoembryonic antigenrelated cell adhesion molecule 1 (CEACAM1), megakaryocyte and plateletinhibitory receptor G6b (G6b-B).

In one embodiment, domains of ITIM receptors LILRB2 (SEQ ID NO: 34),PECAM1 (SEQ ID NO: 38), TLT1 (SEQ ID NO: 43), and CEACAM1 (SEQ ID NO:24) shown in Table 5 may be combined for expression in a megakaryocyteresulting in a CPR in the engineered platelet.

In one embodiment, domains of ITIM receptors may be combined with T cellreceptor domains to form chimeric ITIM receptors which are also referredto as chimeric platelet receptors. These chimeric receptors may becombined for expression in a megakaryocyte resulting in a CPR in theengineered platelet.

TABLE 5 ITIM receptors Sequence Symbol Regions Identifier LILRB2 SignalPeptide (SEQ ID NO: 35); EC Domain 34 (SEQ ID NO: 36); TM Domain (SEQ IDNO: 37); CytoDomain (SEQ ID NO: 52); stop PECAM1 Signal Peptide (SEQ IDNO: 39); EC Domain 38 (SEQ ID NO: 40); TM Domain (SEQ ID NO: 41);CytoDomain (SEQ ID NO: 42); stop TLT1 Signal Peptide (SEQ ID NO: 44); ECDomain 43 (SEQ ID NO: 45); TM Domain (SEQ ID NO: 46); CytoDomain (SEQ IDNO: 47); stop CEACA Signal Peptide (SEQ ID NO: 25); EC Domain 24 M1 (SEQID NO: 26); TM Domain (SEQ ID NO: 27); CytoDomain (SEQ ID NO: 28); stop

SEQ ID NO: 24 is an embodiment of the ITIM receptor of CEACAM1. SEQ IDNO: 25 is an embodiment of the signal peptide of CEACAM1. SEQ ID NO: 26is an embodiment of the extracellular domain of CEACAM1. SEQ ID NO: 27is an embodiment of the transmembrane domain of CEACAM1. SEQ ID NO: 28is an embodiment of the cytoplasmic domain of CEACAM1. SEQ ID NO: 29 isan embodiment of the ITIM receptor of G6b-B. SEQ ID NO: 30 is anembodiment of the signal peptide of G6b-B. SEQ ID NO: 31 is anembodiment of the extracellular domain of G6b-B. SEQ ID NO: 32 is anembodiment of the transmembrane domain of G6b-B. SEQ ID NO: 33 is anembodiment of the cytoplasmic domain of G6b-B. SEQ ID NO: 34 is anembodiment of the ITIM receptor of LILRB2. SEQ ID NO: 35 is anembodiment of the signal peptide of LILRB2. SEQ ID NO: 36 is anembodiment of the extra domain of LILRB2. SEQ ID NO: 37 is an embodimentof the transmembrane domain of LILRB2. SEQ ID NO: 38 is an embodiment ofthe ITIM receptor of PECAM1. SEQ ID NO: 39 is an embodiment of thesignal peptide of PECAM1. SEQ ID NO: 40 is an embodiment of theextracellular domain of PECAM1. SEQ ID NO: 41 is an embodiment of thetransmembrane domain of PECAM1. SEQ ID NO: 42 is an embodiment of thecytoplasmic domain of PECAM1. SEQ ID NO: 43 is an embodiment of the ITIMreceptor of TLT1. SEQ ID NO: 44 is an embodiment of the signal peptideof TLT1. SEQ ID NO: 45 is an embodiment of the extracellular domain ofTLT1. SEQ ID NO: 46 is an embodiment of the transmembrane domain ofTLT1. SEQ ID NO: 47 is an embodiment of the cytoplasmic domain of TLT1.

a. Domains

A CPR may comprise any combination of a signal peptide, an extracellulardomain, a transmembrane domain, a cytoplasmic domain, or linker ortargeting domain.

In one embodiment, a CPR may comprise a signal peptide selected fromTable 6.

TABLE 6 Signal peptides Sequence Identifier Description 2 FCERG SignalPeptide 11 FCGR2 Signal Peptide 16 GPVIA Signal Peptide 25 CEACAM1Signal Peptide 30 G6b-B Signal Peptide 35 LILRB2 Signal Peptide 39PECAM1 Signal Peptide 44 TLT1 Signal Peptide

In one embodiment, a CPR comprises at least one FCERG signal peptide. Asa non-limiting example, the FCERG signal peptide is SEQ ID NO: 2.

In one embodiment, a CPR comprises at least one FCGR2 signal peptide. Asa non-limiting example, the FCGR2 signal peptide is SEQ ID NO: 11.

In one embodiment, a CPR comprises at least one GPVIA signal peptide. Asa non-limiting example, the GPVIA signal peptide is SEQ ID NO: 16.

In one embodiment, a CPR comprises at least one CEACAM1 signal peptide.As a non-limiting example, the CEACAM1 signal peptide is SEQ ID NO: 25.

In one embodiment, a CPR comprises at least one G6b-B signal peptide. Asa non-limiting example, the G6b-B signal peptide is SEQ ID NO: 30.

In one embodiment, a CPR comprises at least one LILRB2 signal peptide.As a non-limiting example, the LILRB2 signal peptide is SEQ ID NO: 35.

In one embodiment, a CPR comprises at least one PECAM1 signal peptide.As a non-limiting example, the PECAM1 signal peptide is SEQ ID NO: 39.

In one embodiment, a CPR comprises at least one TLT1 signal peptide. Asa non-limiting example, the TLT1 signal peptide is SEQ ID NO: 44.

The CPR may include a portion of the signal peptide in Table 6 or asignal peptide known in the art. The portion may be 10-30, 10-15, 10-20,10-25, 15-20, 15-25, 15-30, 20-25, or 20-30, nucleotides of any of thesequences in Table 6 such as, but not limited to, SEQ ID NO: 2, 11, 16,25, 30, 35, 39, and 44. The portion may be 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides ofany of the sequences in Table 7 such as, but not limited to, SEQ ID NO:2, 11, 16, 25, 30, 35, 39, and 44.

A CPR may comprise an extracellular (EC) domain or portion thereofselected from Table 7.

TABLE 7 Extracellular domain Sequence Identifier Description 3 FCERG ECDomain 9 CLEC1 EC Domain 53 CLEC1 EC Domain 12 FCGR2 EC Domain 54 FCGR2EC Domain 17 GPVIA EC Domain 55 GPVIA EC Domain 26 CEACAM1 EC Domain 31G6b-B EC Domain 36 LILRB2 EC Domain 40 PECAM1 EC Domain 45 TLT1 ECDomain

In one embodiment, a CPR comprises at least one FCERG EC domain. As anon-limiting example, the FCERG EC domain is SEQ ID NO: 3.

In one embodiment, a CPR comprises at least one CLEC1 EC domain. As anon-limiting example, the CLEC1 EC domain is SEQ ID NO: 9.

In one embodiment, a CPR comprises at least one CLEC1 EC domain. As anon-limiting example, the CLEC1 EC domain is SEQ ID NO: 53.

In one embodiment, a CPR comprises at least one FCGR2 EC domain. As anon-limiting example, the FCGR2 EC domain is SEQ ID NO: 12.

In one embodiment, a CPR comprises at least one FCGR2 EC domain. As anon-limiting example, the FCGR2 EC domain is SEQ ID NO: 54.

In one embodiment, a CPR comprises at least one GPVIA EC domain. As anon-limiting example, the GPVIA EC domain is SEQ ID NO: 17.

In one embodiment, a CPR comprises at least one GPVIA EC domain. As anon-limiting example, the GPVIA EC domain is SEQ ID NO: 55.

In one embodiment, a CPR comprises at least one CEACAM1 EC domain. As anon-limiting example, the CEACAM1 EC domain is SEQ ID NO: 26.

In one embodiment, a CPR comprises at least one G6b-B EC domain. As anon-limiting example, the G6b-B EC domain is SEQ ID NO: 31.

In one embodiment, a CPR comprises at least one LILRB2 EC domain. As anon-limiting example, the LILRB2 EC domain is SEQ ID NO: 36.

In one embodiment, a CPR comprises at least one PECAM1 EC domain. As anon-limiting example, the PECAM1 EC domain is SEQ ID NO: 40.

In one embodiment, a CPR comprises at least one TLT1 EC domain. As anon-limiting example, the TLT1 EC domain is SEQ ID NO: 45.

The CPR may include a portion of the EC domain in Table 7 or a EC domainknown in the art. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20,15-25, 15-30, 20-25, or 20-30, nucleotides of any of the sequences inTable 7 such as, but not limited to, SEQ ID NO: 3, 9, 12, 17, 26, 31,36, 40, and 45. The portion may be 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides of any ofthe sequences in Table 7 such as, but not limited to, SEQ ID NO: 3, 9,12, 17, 26, 31, 36, 40, and 45.

In one embodiment, CPR may include a portion of the FCERG EC domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 3. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 3.

In one embodiment, CPR may include a portion of the CLEC1 EC domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 9. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 9.

In one embodiment, CPR may include a portion of the FCERG EC domain inTable 7 that is 10-15 nucleotides in length. The portion may be 10, 11,12, 13, 14, or 15 nucleotides of SEQ ID NO: 3.

In one embodiment, CPR may include a portion of the CLEC1 EC domain inTable 7 that is 10-15 nucleotides in length. The portion may be 10, 11,12, 13, 14, or 15 nucleotides of SEQ ID NO: 9 or 53. As a non-limitingexample, a portion of SEQ ID NO: 9 may be SEQ ID NO: 53.

In one embodiment, CPR may include a portion of the CLEC1 EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 53. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 53.

In one embodiment, CPR may include a portion of the FCGR2 EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 12. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 12.

In one embodiment, CPR may include a portion of the FCGR2 EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 54. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 54.

In one embodiment, CPR may include a portion of the GPVIA EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 17. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 17.

In one embodiment, CPR may include a portion of the GPVIA EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 55. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 55.

In one embodiment, CPR may include a portion of the CEACAM1 EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 26. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 26.

In one embodiment, CPR may include a portion of the G6b-B EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 31. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 31.

In one embodiment, CPR may include a portion of the LILRB2 EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 36. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 36.

In one embodiment, CPR may include a portion of the PECAM1 EC Domain inTable 7 The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 40. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 40.

In one embodiment, CPR may include a portion of the TLT1 EC Domain inTable 7. The portion may be 10-30, 10-15, 10-20, 10-25, 15-20, 15-25,15-30, 20-25, or 20-30, nucleotides of SEQ ID NO: 45. The portion may be10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27,28, 29, or 30 nucleotides of SEQ ID NO: 45.

A CPR may comprise a transmembrane (TM) domain selected from Table 8.

TABLE 8 Transmembrane domain Sequence Identifier Description 4 FCERG TMDomain 8 CLEC1 TM Domain 13 FCGR2 TM Domain 18 GPVIA TM Domain 27CEACAM1 TM Domain 32 G6b-B TM Domain 37 LILRB2 TM Domain 41 PECAM1 TMDomain 46 TLT1 TM Domain

In one embodiment, a CPR comprises at least one FCERG TM Domain. As anon-limiting example, the FCERG TM Domain is SEQ ID NO: 4.

In one embodiment, a CPR comprises at least one CLEC1 TM Domain. As anon-limiting example, the CLEC1 TM Domain is SEQ ID NO: 8.

In one embodiment, a CPR comprises at least one FCGR2 TM Domain. As anon-limiting example, the FCGR2 TM Domain is SEQ ID NO: 13.

In one embodiment, a CPR comprises at least one GPVIA TM Domain. As anon-limiting example, the GPVIA TM Domain is SEQ ID NO: 18.

In one embodiment, a CPR comprises at least one CEACAM1 TM Domain. As anon-limiting example, the CEACAM1 TM Domain is SEQ ID NO: 27.

In one embodiment, a CPR comprises at least one G6b-B TM Domain. As anon-limiting example, the G6b-B TM Domain is SEQ ID NO: 32.

In one embodiment, a CPR comprises at least one LILRB2 TM Domain. As anon-limiting example, the LILRB2 TM Domain is SEQ ID NO: 37.

In one embodiment, a CPR comprises at least one PECAM1 TM Domain. As anon-limiting example, the PECAM1 TM Domain is SEQ ID NO: 41.

In one embodiment, a CPR comprises at least one TLT1 TM Domain. As anon-limiting example, the TLT1 TM Domain is SEQ ID NO: 46.

A CPR may comprise a cytoplasm (cyto) domain selected from Table 9.

TABLE 9 Cytoplasm domain Sequence Identifier Description 5 FCERGCytoDomain 7 CLEC1 CytoDomain 14 FCGR2 CytoDomain 19 GPVIA CytoDomain 28CEACAM1 CytoDomain 33 G6b-B CytoDomain 52 LILRB2 CytoDomain 42 PECAM1CytoDomain 47 TLT1 CytoDomain

In one embodiment, a CPR comprises at least one FCERG CytoDomain. As anon-limiting example, the FCERG CytoDomain is SEQ ID NO: 5.

In one embodiment, a CPR comprises at least one CLEC1 CytoDomain. As anon-limiting example, the CLEC1 CytoDomain is SEQ ID NO: 7.

In one embodiment, a CPR comprises at least one FCGR2 CytoDomain. As anon-limiting example, the FCGR2 CytoDomain is SEQ ID NO: 14.

In one embodiment, a CPR comprises at least one GPVIA CytoDomain. As anon-limiting example, the GPVIA CytoDomain is SEQ ID NO: 19.

In one embodiment, a CPR comprises at least one CEACAM1 CytoDomain. As anon-limiting example, the CEACAM1 CytoDomain is SEQ ID NO: 28.

In one embodiment, a CPR comprises at least one G6b-B CytoDomain. As anon-limiting example, the G6b-B CytoDomain is SEQ ID NO: 33.

In one embodiment, a CPR comprises at least one LILRB2 CytoDomain. As anon-limiting example, the LILRB2 CytoDomain is SEQ ID NO: 52.

In one embodiment, a CPR comprises at least one PECAM1 CytoDomain. As anon-limiting example, the PECAM1 CytoDomain is SEQ ID NO: 42.

In one embodiment, a CPR comprises at least one TLT1 CytoDomain. As anon-limiting example, the TLT1 CytoDomain is SEQ ID NO: 47.

Genes encoding fusion peptides, targeting domain, or linking protein maybe added to the genome of the megakaryocyte as shown in Table 10, suchas the L chain of variable fragment 1 (Fv1_Lchain) with a nucleic acidsequence of SEQ ID NO: 48 or an improved linker from a single-chainvariable fragment with reduced aggregation and enhanced proteolyticstability (Whitlow_linker) with a nucleic acid sequence of SEQ ID NO:49. Alternatively, at least a portion of an antibody may be added to thegenome of the megakaryocyte for expression in a resulting platelet, suchas a kappa light chain of an anti-human B cell CD19 antibody(F1HChain_CD19FMC63) with a nucleic acid sequence of SEQ ID NO: 50 and amodified IGg4 hinge region with a nucleic acid sequence of SEQ ID NO: 51also shown in Table 10.

In some embodiments, a CPR comprises at least one domain selected fromTables 4-9 and a linker and/or targeting domains selected from Table 10.

TABLE 10 Linkers and Targeting Domains Sequence Identifier Description48 Fv1_Lchain (Light Chain) 49 Whitlow_linker (Linker) 50Fv1HChain_CD19FMC63 (Heavy Chain) 51 Modified_hinge_IGg4 (Hinge)

In one embodiment, the CPR has a domain, having at least 95% identity toany of the sequences of Tables 4-10, including, SEQ ID NO: 1-55. In oneembodiment, the CPR has a domain having 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, or 100% identity to one or more of the sequences of Tables4-10, including, SEQ ID NO: 1-55.

In one embodiment, the CPR has at least one signal peptide, having atleast 95% identity to any of the sequences of Table 6, including, SEQ IDNO: 2, 11, 16, 25, 30, 35, 39, and 44. In one embodiment, the CPR has atleast one signal peptide domain having 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, or 100% identity to one or more of the sequences of Table 6,including, SEQ ID NO: 2, 11, 16, 25, 30, 35, 39, and 44.

In one embodiment, the CPR has at least one extracellular domain, havingat least 95% identity to any of the sequences of Table 7, including, SEQID NO: 3, 9, 53, 12, 54, 17, 55, 26, 31, 36, 40, or 45. In oneembodiment, the CPR has at least one extracellular domain having 50%,51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%,65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%,79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to one or more ofthe sequences of Table 7, including, SEQ ID NO: 3, 9, 53, 12, 54, 17,55, 26, 31, 36, 40, or 45.

In one embodiment, the CPR has at least one transmembrane domain, havingat least 95% identity to any of the sequences of Table 8, including, SEQID NO: 4, 8, 13, 18, 27, 32, 37, 41, or 46. In one embodiment, the CPRhas at least one transmembrane domain having 50%, 51%, 52%, 53%, 54%,55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%,69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90° %, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99%, or 100% identity to one or more of the sequences of Table8, including, SEQ ID NO: 4, 8, 13, 18, 27, 32, 37, 41, or 46.

In one embodiment, the CPR has at least one cytoplasmic domain, havingat least 95% identity to any of the sequences of Table 9, including, SEQID NO: 5, 7, 14, 19, 28, 33, 52, 42, or 47. In one embodiment, the CPRhas at least one cytoplasmic domain having 50%, 51%, 52%, 53%, 54%, 55%,56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%,70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%,84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, 99%, or 100% identity to one or more of the sequences of Table 9,including, SEQ ID NO: 5, 7, 14, 19, 28, 33, 52, 42, or 47.

In one embodiment, the CPR has at least one linker or targeting domain,having at least 95% identity to any of the sequences of Table 10,including, SEQ ID NO: 48, 49, 50, or 51. In one embodiment, the CPR hasat least one linker or targeting domain having 50%, 51%, 52%, 53%, 54%,55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%,69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99%, or 100% identity to one or more of the sequences of Table10, including, SEQ ID NO: 48, 49, 50, or 51.

In one embodiment, the CPR comprises a signal peptide, extracellulardomain, a transmembrane domain and a cytoplasm domain.

In one embodiment, the CPR comprises an extracellular domain, atransmembrane domain and a cytoplasm domain.

b. Domain Swapping

In one embodiment, any of the domains in the ITAM and/or ITIM receptorsin Tables 4 and 5, respectfully, may be replaced with domains from otherITAM and/or ITIM receptors.

In one embodiment, the EC domain in the ITAM receptors in Table 4 may bereplaced with domains from other ITAM and/or ITIM receptors. Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aCLEC1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aFCGR2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aGPVIA EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aLILRB2 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aPECAM1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCERG ITAM receptor may be replaced by aCEACAM1 EC domain or a portion thereof in order to create a CPR.

For example, the TM domain in the FCERG ITAM receptor may be replaced bya CLEC1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCERG ITAM receptor may be replaced by aFCGR2 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCERG ITAM receptor may be replaced by aGPVIA TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the FCERG ITAM receptor may be replaced by aLILRB2 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the FCERG ITAM receptor may be replaced by aPECAM1 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the FCERG ITAM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCERG ITAM receptor may be replaced by aCEACAM1 TM domain or a portion thereof in order to create a CPR.

For example, the signal peptide in the FCERG ITAM receptor may bereplaced by a CLEC1 signal peptide or a portion thereof in order tocreate a CPR For example, the signal peptide in the FCERG ITAM receptormay be replaced by a FCGR2 signal peptide or a portion thereof in orderto create a CPR For example, the signal peptide in the FCERG ITAMreceptor may be replaced by a GPVIA signal peptide or a portion thereofin order to create a CPR. For example, the signal peptide in the FCERGITAM receptor may be replaced by a LILRB2 signal peptide or a portionthereof in order to create a CPR For example, the signal peptide in theFCERG ITAM receptor may be replaced by a PECAM1 signal peptide or aportion thereof in order to create a CPR. For example, the signalpeptide in the FCERG ITAM receptor may be replaced by a TLT1 signalpeptide or a portion thereof in order to create a CPR. For example, thesignal peptide in the FCERG ITAM receptor may be replaced by a CEACAM1signal peptide or a portion thereof in order to create a CPR.

For example, the cytodomain in the FCERG ITAM receptor may be replacedby a CLEC1 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the FCERG ITAM receptor may be replaced by aFCGR2 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCERG ITAM receptor may be replaced by aGPVIA cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCERG ITAM receptor may be replaced by aLILRB2 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCERG ITAM receptor may be replaced by aPECAM1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCERG ITAM receptor may be replaced by aTLT1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCERG ITAM receptor may be replaced by aCEACAM1 cytodomain or a portion thereof in order to create a CPR.

For example, the EC domain in the CLEC1 ITAM receptor may be replaced bya FCGR2 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the CLEC1 ITAM receptor may be replaced by aGPVIA EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CLEC1 ITAM receptor may be replaced by aLILRB2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CLEC1 ITAM receptor may be replaced by aPECAM1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the CLEC1 ITAM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CLEC1 ITAM receptor may be replaced by aCEACAM1 EC domain or a portion thereof in order to create a CPR

For example, the TM domain in the CLEC1 ITAM receptor may be replaced bya FCERG TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the CLEC1 ITAM receptor may be replaced by aFCGR2 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CLEC1 ITAM receptor may be replaced by aGPVIA TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the CLEC1 ITAM receptor may be replaced by aLILRB2 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CLEC1 ITAM receptor may be replaced by aPECAM1 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the CLEC1 ITAM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CLEC1 ITAM receptor may be replaced by aCEACAM1 TM domain or a portion thereof in order to create a CPR.

For example, the signal peptide in the CLEC1 ITAM receptor may bereplaced by a FCERG signal peptide or a portion thereof in order tocreate a CPR For example, the signal peptide in the CLEC1 ITAM receptormay be replaced by a FCGR2 signal peptide or a portion thereof in orderto create a CPR. For example, the signal peptide in the CLEC1 ITAMreceptor may be replaced by a GPVIA signal peptide or a portion thereofin order to create a CPR. For example, the signal peptide in the CLEC1ITAM receptor may be replaced by a LILRB2 signal peptide or a portionthereof in order to create a CPR. For example, the signal peptide in theCLEC1 ITAM receptor may be replaced by a PECAM1 signal peptide or aportion thereof in order to create a CPR. For example, the signalpeptide in the CLEC1 ITAM receptor may be replaced by a TLT1 signalpeptide or a portion thereof in order to create a CPR. For example, thesignal peptide in the CLEC1 ITAM receptor may be replaced by a CEACAM1signal peptide or a portion thereof in order to create a CPR.

For example, the cytodomain in the CLEC1 ITAM receptor may be replacedby a FCERG cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the CLEC1 ITAM receptor may be replaced by aFCGR2 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CLEC1 ITAM receptor may be replaced by aGPVIA cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CLEC1 ITAM receptor may be replaced by aLILRB2 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the CLEC1 ITAM receptor may be replaced by aPECAM1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CLEC1 ITAM receptor may be replaced by aTLT1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CLEC1 ITAM receptor may be replaced by aCEACAM1 cytodomain or a portion thereof in order to create a CPR

For example, the EC domain in the FCGR2 ITAM receptor may be replaced bya CLEC1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the FCGR2 ITAM receptor may be replaced by aGPVIA EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCGR2 ITAM receptor may be replaced by aLILRB2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the FCGR2 ITAM receptor may be replaced by aPECAM1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the FCGR2 ITAM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the FCGR2 ITAM receptor may be replaced by aCEACAM1 EC domain or a portion thereof in order to create a CPR.

For example, the TM domain in the FCGR2 ITAM receptor may be replaced bya FCERG TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCGR2 ITAM receptor may be replaced by aCLEC1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCGR2 ITAM receptor may be replaced by aGPVIA TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCGR2 ITAM receptor may be replaced by aLILRB2 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the FCGR2 ITAM receptor may be replaced by aPECAM1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCGR2 ITAM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the FCGR2 ITAM receptor may be replaced by aCEACAM1 TM domain or a portion thereof in order to create a CPR

For example, the signal peptide in the FCGR2 ITAM receptor may bereplaced by a FCERG signal peptide or a portion thereof in order tocreate a CPR For example, the signal peptide in the FCGR2 ITAM receptormay be replaced by a CLEC1 signal peptide or a portion thereof in orderto create a CPR For example, the signal peptide in the FCGR2 ITAMreceptor may be replaced by a GPVIA signal peptide or a portion thereofin order to create a CPR For example, the signal peptide in the FCGR2ITAM receptor may be replaced by a LILRB2 signal peptide or a portionthereof in order to create a CPR For example, the signal peptide in theFCGR2 ITAM receptor may be replaced by a PECAM1 signal peptide or aportion thereof in order to create a CPR For example, the signal peptidein the FCGR2 ITAM receptor may be replaced by a TLT1 signal peptide or aportion thereof in order to create a CPR For example, the signal peptidein the FCGR2 ITAM receptor may be replaced by a CEACAM1 signal peptideor a portion thereof in order to create a CPR

For example, the cytodomain in the FCGR2 ITAM receptor may be replacedby a FCERG cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the FCGR2 ITAM receptor may be replaced by aCLEC1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCGR2 ITAM receptor may be replaced by aGPVIA cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCGR2 ITAM receptor may be replaced by aLILRB2 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the FCGR2 ITAM receptor may be replaced by aPECAM1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCGR2 ITAM receptor may be replaced by aTLT1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the FCGR2 ITAM receptor may be replaced by aCEACAM1 cytodomain or a portion thereof in order to create a CPR.

For example, the EC domain in the GPVIA ITAM receptor may be replaced bya CLEC1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the GPVIA ITAM receptor may be replaced by aFCGR2 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the GPVIA ITAM receptor may be replaced by aLILRB2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the GPVIA ITAM receptor may be replaced by aPECAM1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the GPVIA ITAM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the GPVIA ITAM receptor may be replaced by aCEACAM1 EC domain or a portion thereof in order to create a CPR.

For example, the TM domain in the GPVIA ITAM receptor may be replaced bya FCERG TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the GPVIA ITAM receptor may be replaced by aCLEC1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the GPVIA ITAM receptor may be replaced by aFCGR2 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the GPVIA ITAM receptor may be replaced by aLILRB2 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the GPVIA ITAM receptor may be replaced by aPECAM1 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the GPVIA ITAM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the GPVIA ITAM receptor may be replaced by aCEACAM1 TM domain or a portion thereof in order to create a CPR.

For example, the signal peptide in the GPVIA ITAM receptor may bereplaced by a FCERG signal peptide or a portion thereof in order tocreate a CPR For example, the signal peptide in the GPVIA ITAM receptormay be replaced by a CLEC1 signal peptide or a portion thereof in orderto create a CPR For example, the signal peptide in the GPVIA ITAMreceptor may be replaced by a FCGR2 signal peptide or a portion thereofin order to create a CPR. For example, the signal peptide in the GPVIAITAM receptor may be replaced by a LILRB2 signal peptide or a portionthereof in order to create a CPR For example, the signal peptide in theGPVIA ITAM receptor may be replaced by a PECAM1 signal peptide or aportion thereof in order to create a CPR. For example, the signalpeptide in the GPVIA ITAM receptor may be replaced by a TLT1 signalpeptide or a portion thereof in order to create a CPR. For example, thesignal peptide in the GPVIA ITAM receptor may be replaced by a CEACAM1signal peptide or a portion thereof in order to create a CPR.

For example, the cytodomain in the GPVIA ITAM receptor may be replacedby a FCERG cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the GPVIA ITAM receptor may be replaced by aCLEC1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the GPVIA ITAM receptor may be replaced by aFCGR2 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the GPVIA ITAM receptor may be replaced by aLILRB2 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the GPVIA ITAM receptor may be replaced by aPECAM1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the GPVIA ITAM receptor may be replaced by aTLT1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the GPVIA ITAM receptor may be replaced by aCEACAM1 cytodomain or a portion thereof in order to create a CPR.

In one embodiment, the EC domain in the ITIM receptors in Table 5 may bereplaced with domains from other ITAM and/or ITIM receptors.

For example, the EC domain in the LILRB2 ITIM receptor may be replacedby a CLEC1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the LILRB2 ITIM receptor may be replaced by aGPVIA EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the LILRB2 ITIM receptor may be replaced by aFCGR2 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the LILRB2 ITIM receptor may be replaced by aPECAM1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the LILRB2 ITIM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the LILRB2 ITIM receptor may be replaced by aCEACAM1 EC domain or a portion thereof in order to create a CPR.

For example, the TM domain in the LILRB2 ITIM receptor may be replacedby a FCERG TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the LILRB2 ITIM receptor may be replaced by aCLEC1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the LILRB2 ITIM receptor may be replaced by aGPVIA TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the LILRB2 ITIM receptor may be replaced by aFCGR2 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the LILRB2 ITIM receptor may be replaced by aPECAM1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the LILRB2 ITIM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the LILRB2 ITIM receptor may be replaced by aCEACAM1 TM domain or a portion thereof in order to create a CPR

For example, the signal peptide in the LILRB2 ITIM receptor may bereplaced by a FCERG signal peptide or a portion thereof in order tocreate a CPR For example, the signal peptide in the LILRB2 ITIM receptormay be replaced by a CLEC1 signal peptide or a portion thereof in orderto create a CPR For example, the signal peptide in the LILRB2 ITIMreceptor may be replaced by a GPVIA signal peptide or a portion thereofin order to create a CPR. For example, the signal peptide in the LILRB2ITIM receptor may be replaced by a FCGR2 signal peptide or a portionthereof in order to create a CPR For example, the signal peptide in theLILRB2 ITIM receptor may be replaced by a PECAM1 signal peptide or aportion thereof in order to create a CPR. For example, the signalpeptide in the LILRB2 ITIM receptor may be replaced by a TLT1 signalpeptide or a portion thereof in order to create a CPR For example, thesignal peptide in the LILRB2 ITIM receptor may be replaced by a CEACAM1signal peptide or a portion thereof in order to create a CPR

For example, the cytodomain in the LILRB2 ITIM receptor may be replacedby a FCERG cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the LILRB2 ITIM receptor may be replaced by aCLEC1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the LILRB2 ITIM receptor may be replaced by aGPVIA cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the LILRB2 ITIM receptor may be replaced by aFCGR2 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the LILRB2 ITIM receptor may be replaced by aPECAM1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the LILRB2 ITIM receptor may be replaced by aTLT1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the LILRB2 ITIM receptor may be replaced by aCEACAM1 cytodomain or a portion thereof in order to create a CPR. Forexample, the EC domain in the PECAM1 ITIM receptor may be replaced by aCLEC1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the PECAM1 ITIM receptor may be replaced by aGPVIA EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the PECAM1 ITIM receptor may be replaced by aFCGR2 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the PECAM1 ITIM receptor may be replaced by aLILRB2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the PECAM1 ITIM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the PECAM1 ITIM receptor may be replaced by aCEACAM1 EC domain or a portion thereof in order to create a CPR

For example, the TM domain in the PECAM1 ITIM receptor may be replacedby a FCERG TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the PECAM1 ITIM receptor may be replaced by aCLEC1 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the PECAM1 ITIM receptor may be replaced by aGPVIA TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the PECAM1 ITIM receptor may be replaced by aFCGR2 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the PECAM1 ITIM receptor may be replaced by aLILRB2 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the PECAM1 ITIM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the PECAM1 ITIM receptor may be replaced by aCEACAM1 TM domain or a portion thereof in order to create a CPR.

For example, the signal peptide in the PECAM1 ITIM receptor may bereplaced by a FCERG signal peptide or a portion thereof in order tocreate a CPR For example, the signal peptide in the PECAM1 ITIM receptormay be replaced by a CLEC1 signal peptide or a portion thereof in orderto create a CPR For example, the signal peptide in the PECAM1 ITIMreceptor may be replaced by a GPVIA signal peptide or a portion thereofin order to create a CPR. For example, the signal peptide in the PECAM1ITIM receptor may be replaced by a FCGR2 signal peptide or a portionthereof in order to create a CPR For example, the signal peptide in thePECAM1 ITIM receptor may be replaced by a LILRB2 signal peptide or aportion thereof in order to create a CPR For example, the signal peptidein the PECAM1 ITIM receptor may be replaced by a TLT1 signal peptide ora portion thereof in order to create a CPR For example, the signalpeptide in the PECAM1 ITIM receptor may be replaced by a CEACAM1 signalpeptide or a portion thereof in order to create a CPR.

For example, the cytodomain in the PECAM1 ITIM receptor may be replacedby a FCERG cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the PECAM1 ITIM receptor may be replaced by aCLEC1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the PECAM1 ITIM receptor may be replaced by aGPVIA cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the PECAM1 ITIM receptor may be replaced by aFCGR2 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the PECAM1 ITIM receptor may be replaced by aLILRB2 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the PECAM1 ITIM receptor may be replaced by aTLT1 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya CEACAM1 cytodomain or a portion thereof in order to create a CPR.

For example, the EC domain in the CEACAM1 ITIM receptor may be replacedby a CLEC1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CEACAM1 ITIM receptor may be replaced by aGPVIA EC domain or a portion thereof in order to create a CPR. Forexample, the EC domain in the CEACAM1 ITIM receptor may be replaced by aFCGR2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CEACAM1 ITIM receptor may be replaced by aLILRB2 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CEACAM1 ITIM receptor may be replaced by aPECAM1 EC domain or a portion thereof in order to create a CPR Forexample, the EC domain in the CEACAM1 ITIM receptor may be replaced by aTLT1 EC domain or a portion thereof in order to create a CPR

For example, the TM domain in the CEACAM1 ITIM receptor may be replacedby a FCERG TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CEACAM1 ITIM receptor may be replaced by aCLEC1 TM domain or a portion thereof in order to create a CPR Forexample, the TM domain in the CEACAM1 ITIM receptor may be replaced by aGPVIA TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CEACAM1 ITIM receptor may be replaced by aFCGR2 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CEACAM1 ITIM receptor may be replaced by aLILRB2 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CEACAM1 ITIM receptor may be replaced by aPECAM1 TM domain or a portion thereof in order to create a CPR. Forexample, the TM domain in the CEACAM1 ITIM receptor may be replaced by aTLT1 TM domain or a portion thereof in order to create a CPR.

For example, the signal peptide in the CEACAM1 ITIM receptor may bereplaced by a FCERG signal peptide or a portion thereof in order tocreate a CPR. For example, the signal peptide in the CEACAM1 ITIMreceptor may be replaced by a CLEC1 signal peptide or a portion thereofin order to create a CPR For example, the signal peptide in the CEACAM1ITIM receptor may be replaced by a GPVIA signal peptide or a portionthereof in order to create a CPR For example, the signal peptide in theCEACAM1 ITIM receptor may be replaced by a FCGR2 signal peptide or aportion thereof in order to create a CPR For example, the signal peptidein the CEACAM1 ITIM receptor may be replaced by a LILRB2 signal peptideor a portion thereof in order to create a CPR For example, the signalpeptide in the CEACAM1 ITIM receptor may be replaced by a PECAM1 signalpeptide or a portion thereof in order to create a CPR For example, thesignal peptide in the CEACAM1 ITIM receptor may be replaced by a TLT1signal peptide or a portion thereof in order to create a CPR.

For example, the cytodomain in the CEACAM1 ITIM receptor may be replacedby a FCERG cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya CLEC1 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya GPVIA cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya FCGR2 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya LILRB2 cytodomain or a portion thereof in order to create a CPR. Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya PECAM1 cytodomain or a portion thereof in order to create a CPR Forexample, the cytodomain in the CEACAM1 ITIM receptor may be replaced bya TLT1 cytodomain or a portion thereof in order to create a CPR

In some embodiments, the signal peptide, EC domain, TM domain, orcytodomain of an ITIM or ITAM reception may be replaced by a portion ofa domain from a different receptor. The portion may have a length withinthe range of 10-30, 10-15, 10-20, 10-25, 15-20, 15-25, 15-30, 20-25, or20-30 nucleotides. The portion may be 10, 11, 12, 13, 14, 15, 16, 17,18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides of anyof the sequences in Tables 6-10 herein, for example, but not limited to,SEQ ID NO: 1-55. In certain embodiments, an EC domain from a CLEC1 ITAMreceptor is SEQ ID NO: 53. In certain embodiments, an EC domain from aFCGR2 ITAM receptor is SEQ ID NO: 54. In certain embodiments, an ECdomain from a GPVIA ITAM receptor is SEQ ID NO: 55.

2. Antibodies or Fragments Thereof

The CPR may include a region comprising any ligand or fragment thereofor epitope that recognizes and binds to a target, for example, aneoantigen on tumor cells, with high specificity and affinity. Theregion of the CPR may be an antibody or variants thereof or a variant orfragment thereof that can bind to the target on tumor cells such as atumor specific antigen (TSA), or recognition components such as a linkedcytokine that leads to recognition of target cells bearing the cytokinereceptor, or a natural ligand of a receptor or fragment thereof. In someembodiments, the CPR of the engineered platelets described herein may bea scFv. An antibody, variant, or fragment thereof can be generated usingroutine recombinant DNA technology techniques known in the art.

In some embodiments, the engineered platelets comprise a CPR with aregion including an antibody or an antibody fragment to bind a target ofinterest. For example, the CPR may include a variable heavy chain domainof an antibody. For example, the CPR may include a variable light chaindomain of an antibody. Alternatively, the CPR of the engineeredplatelets may include a kappa light chain or a fragment thereoftargeting CD19.

In some embodiments, the antibody or the antibody fragment thereof ischosen from Table 11. The antibodies are listed with their DrugBankidentifier (DB ID).

TABLE 11 Antibodies for use in chimeric receptors Antibody IGMT/ INN(International INN (Given Name) mAb-DB ID Nonproprietary Name) NumberAbagovomab 111 abagovomab 8659 Abciximab 146 abciximab 7200 Abituzumab489 abituzumab 9509 Abrezekimab 804 abrezekimab 10745 Abrilumab 495abrilumab 9921 Actoxumab 410 actoxumab 9568 Adalimumab 165 adalimumab7860 Adecatumumab 164 adecatumumab 8449 Atidortoxumab 763 atidortoxumab10638 Aducanumab 479 aducanumab 9838 Afasevikumab 576 afasevikumab 10178Afelimomab 128 afelimomab 7340 Alemtuzumab 11 alemtuzumab 8005Alirocumab 412 alirocumab 9620 Amatuximab 64 amatuximab 9343 Anatumomabmafenatox 287 anatumomab mafenatox 7655 Andecaliximab 533 andecaliximab10035 Anetumab ravtansine 471 anetumab ravtansine 9788 Anifrolumab 474anifrolumab 9800 Anrukinzumab 231 anrukinzumab 8942 Apolizumab 18apolizumab 8210 Aprutumab ixadotin 656 aprutumab ixadotin 10383Arcitumomab 113 arcitumomab 7417 Ascrinvacumab 561 ascrinvacumab 10106Aselizumab 272 aselizumab 8291 Atezolizumab 526 atezolizumab 9814Atinumab 358 atinumab 9336 Atorelimumab 308 atorolimumab 7647 Avelumab512 avelumab 10062 Azintuxizumab vedotin 721 azintuxizumab vedotin 10499Bapineuzumab 252 bapineuzumab 8624 Basiliximab 148 basiliximab 7578Bavituximab 149 bavituximab 8734 Bectumomab 306 bectumomab 7514Begelomab 503 begelomab 9959 Belantamab mafodotin 784 belantamabmafodotin 10754 Belimumab 266 belimumab 8381 Bemarituzumab 770bemarituzumab 10681 Benralizumab 334 benralizumab 9233 Berlimatoxumab764 berlimatoxumab 10639 Bersanlimab 800 bersanlimab 10709 Bertilimumab170 bertilimumab 8332 Besilesomab 258 besilesomab 8451 Bevacizumab 24bevacizumab 8017 Bezlotoxumab 411 bezlotoxumab 9608 Biciromab 131biciromab 6867 Bimagrumab 456 bimagrumab 9711 Bimekizumab 486bimekizumab 9878 Birtamimab 619 birtamimab 10198 Bleselumab 563bleselumab 10114 Blinatumomab 101 blinatumomab 9028 Blontuvetmab 593blontuvetmab 10194 Blosozumab 375 blosozumab 9440 Bococizumab 485bococizumab 9840 Brazikumab 664 brazikumab 10425 Brentuximab vedotin 324brentuximab vedotin 9144 Briakinumab 162 briakinumab 9153 Brodalumab 376brodalumab 9475 Brolucizumab 536 brolucizumab 10053 Brontictuzumab 492brontictuzumab 9982 Burosumab 647 burosumab 10301 Cabiralizumab 587cabiralizumab 10121 Camidanlumab tesirine 775 camidanlumab tesirine10592 Camrelizumab 659 camrelizumab 10400 Canakinumab 244 canakinumab8836 Cantuzumab mertansine 52 cantuzumab mertansine 8223 Cantuzumabravtansine 387 cantuzumab ravtansine 9441 Caplacizumab 401 caplacizumab9511 Carlumab 359 carlumab 9372 Carotuximab 605 carotuximab 10244Catumaxomab 218 Catumaxomab 8406 Cedelizumab 78 cedelizumab 7567Cemiplimab 846 cemiplimab 10691 Cergutuzumab amunaleukin 555cergutuzumab amunaleukin 10080 Certolizumab pegol 242 certolizumab pegol8448 Cetrelimab 809 cetrelimab 10757 Cetuximab 151 cetuximab 7906Cibisatamab 795 cibisatamab 10636 Citatuzumab bogatox 236 citatuzumabbogatox 9046 Cixutumumab 290 cixutumumab 9099 Clazakizumab 414clazakizumab 9599 Clenoliximab 152 clenoliximab 7615 Clivatuzumabtetraxetan 560 clivatuzumab tetraxetan 10103 Codrituzumab 466codrituzumab 9759 Cofetuzumab pelidotin 777 cofetuzumab pelidotin 10674Coltuximab ravtansine 490 coltuximab ravtansine 9558 Conatumumab 224conatumumab 9029 Concizumab 447 concizumab 9636 Cosfroviximab 726cosfroviximab 10535 Crenezumab 378 crenezumab 9482 Crizanlizumab 667crizanlizumab 10316 Crotedumab 595 crotedumab 10196 Cusatuzumab 792cusatuzumab 10558 Dacetuzumab 232 dacetuzumab 8959 Daclizumab 36daclizumab 7164 Dalotuzumab 333 dalotuzumab 9200 Dapirolizumab pegol 481dapirolizumab pegol 9869 Daratumumab 301 daratumumab 9128 Dectrekumab537 dectrekumab 10059 Demcizumab 415 demcizumab 9572 Denintuzumabmafodotin 493 denintuzumab mafedotin 9886 Denosumab 249 denosumab 8653Depatuxizumab mafodotin 645 depatuxizumab mafedotin 10263 Detumomab 313detumomab 7156 Dezamizumab 652 dezamizumab 10364 Dinutuximab 464dinutuximab 9754 Diridavumab 494 diridavumab 9922 Domagrozumab 610domagrozumab 10286 Dorlimomab aritox 207 dorlimomab aritox 6516Dostarlimab 849 dostarlimab 10787 Drozitumab 348 drozitumab 9255Duligotuzumab 416 duligotuzumab 9646 Dupilumab 449 dupilumab 9669Durvalumab 528 durvalumab 10010 Dusigitumab 451 dusigitumab 9679Duvortuxizumab 640 duvortuxizumab 10506 Ecromeximab 274 ecromeximab 8239Eculizumab 37 eculizumab 8231 Edobacomab 118 edobacomab 7056 Edrecolomab119 edrecolomab 7471 Efalizumab 38 efalizumab 8122 Efungumab 173efungumab 8658 Eldelumab 463 eldelumab 9746 Elezanumab 650 elezanumab10344 Elgemtumab 534 elgemtumab 10041 Elotuzumab 291 elotuzumab 9074Elsilimomab 268 elsilimomab 8371 Emactuzumab 501 emactuzumab 9951Emapalumab 666 emapalumab 10319 Emibetuzumab 496 emibetuzumab 9932Emicizumab 564 emicizumab 10115 Enapotamab vedotin 814 enapotamabvedotin 10769 Enavatuzumab 360 enavatuzumab 9354 Enfortumab vedotin 476enfortumab vedotin 9821 Enlimomab pegol 317 enlimomab pegol 7525Enoblituzumab 590 enoblituzumab 10165 Enokizumab 361 enokizumab 9262Enoticumab 417 enoticumab 9575 Ensituximab 349 ensituximab 9300Epitumomab cituxetan 269 epitumomab cituxetan 8372 Epratuzumab 40epratuzumab 7920 Eptinezumab 648 eptinezumab 10308 Erenumab 618 erenumab10296 Erlizumab 280 erlizumab 8076 Ertumaxomab 219 ertumaxomab 8407Etaricizumab 240 etaracizumab 8862 Etigilimab 803 etigilimab 10742Etrolizumab 362 etrolizumab 9290 Evinacumab 529 evinacumab 10013Evolocumab 448 evolocumab 9643 Exbivirumab 262 exbivirumab 8536Faralimomab 305 faralimomab 7496 Faricimab 793 faricimab 10563Farletuzumab 292 farletuzumab 9067 Fasinumab 418 fasinumab 9589Felvizumab 86 felvizumab 7623 Fezakinumab 323 fezakinumab 9137Ficlatuzumab 379 ficlatuzumab 9465 Figitumumab 293 figitumumab 9085Firivumab 506 firivumab 9975 Flanvotumab 403 flanvotumab 9520 Fletikumab480 fletikumab 9876 Flotetuzumab 679 flotetuzumab 10569 Fontolizumab 43fontolizumab 8264 Foralumab 350 foralumab 9309 Foravirumab 90foravirumab 9053 Fremanezumab 646 fremanezumab 10299 Fresolimumab 174fresolimumab 9158 Frovocimab 850 frovocimab 10859 Frunevetmab 717frunevetmab 10440 Fulranumab 363 fulranumab 9373 Futuximab 419 futuximab9612 Galcanezumab 609 galcanezumab 10277 Galiximab 154 galiximab 8339Gancotamab 865 gancotamab 10562 Ganitumab 168 ganitumab 9323Gantenerumab 223 gantenerumab 8894 Gatipotuzumab 740 gatipotuzumab 10336Gavilimomab 112 gavilimomab 8060 Gedivumab 719 gedivumab 10459Gemtuzumab ozogamicin 649 gemtuzumab ozogamicin 10315 Gevokizumab 102gevokizumab 9310 Gilvetmab 725 gilvetmab 10528 Gimsilumab 756 gimsilumab10534 Girentuximab 328 girentuximab 9116 Glembatumumab vedotin 565glembatumumab vedotin 10123 Golimumab 175 golimumab 8497 Gosuranemab 851gosuranemab 10663 Guselkumab 468 guselkumab 9774 Ianalumab 758 ianalumab10580 Ibalizumab 241 ibalizumab 8818 Ibritumomab tiuxetan 122ibritumomab tiuxetan 7873 Icrumcumab 365 icrucumab 9370 Idarucizumab 462idarucizumab 9698 Ifabotuzumab 572 ifabotuzumab 10149 Igovomab 123igovomab 7433 Iladatuzumab vedotin 774 iladatuzumab vedotin 10647Imalumab 504 imalumab 9961 Imaprelimab 808 imaprelimab 10753 Imciromab124 imciromab 6605 Imgatuzumab 420 imgatuzumab 9598 Inclacumab 402.inclacumab 9512 Indatuximab ravtansine 389 indatuximab ravtansine 9486Indusatumab vedotin 532 indusatumab vedotin 10033 Inebilizumab 553inebilizumab 9985 Infliximab 156 infliximab 7602 intetumumab 321intetumumab 9134 inolimomab 126 inolimomab 7253 inotuzumab ozogamicin259 inotuzumab ozogamicin 8574 Ipilimumab 180 ipilimumab 8568 Iratumumab250 iratumumab 8713 Isatuximab 539 isatuximab 10068 Iscalimab 799iscalimab 10707 Istiratumab 545 istiratumab 10431 Itolizumab 351itolizumab 9321 Ixekizumab 380 ixekizumab 9467 Keliximab 157 keliximab7560 Labetuzumab 62 labetuzumab 8127 Lacnotuzumab 724 lacnotuzumab 10524Ladiratuzumab vedotin 773 ladiratuzumab vedotin 10625 Lampalizumab 421lampalizumab 9578 Lanadelumab 607 lanadelumab 10265 Landogrozumab 578landogrozumab 10188 Laprituximab emtansine 600 laprituximab emtansine10236 Larcaviximab 728 larcaviximab 10537 Lebrikizumab 325 lebrikizumab9165 Lemalesomab 281 lemalesomab 8046 Lenvervimab 805 lenvervimab 10746Lenzilumab 505 lenzilumab 9965 Lerdelimumab 182 lerdelimumab 7882Leronlimab 807 leronlimab 10751 Lesofavumab 718 lesofavumab 10458Letolizumab 715 letolizumab 10436 Lexatumumab 183 lexatumumab 8753Libivirumab 263 libivirumab 8481 Lifastuzumab vedotin 478 lifastuzumabvedotin 9835 Ligelizumab 422 ligelizumab 9653 Loncastuximab tesirine 749loncastuximab tesirine 10586 Losatuxizumab vedotin 712 losatuxizumabvedotin 10342 Lintuzumab 53 lintuzumab 7580 Lirilumab 423 lirilumab 9415Lodelcizumab 461 lodelcizumab 9733 Lokivetmab 527 lokivetmab 10002Lorvotuzumab mertansine 58 lorvotuzumab mertansine 9299 Lucatumuntab 176lucatumumab 8887 Lulizumab pegol 499 lulizumab pegol 9940 Lumiliximab158 lumiliximab 8443 Lumretuzumab 502 lumretuzumab 9952 Lupartumabamadotin 665 lupartumab amadotin 10257 Lutikizumab 651 lutikizumab 10347Mapatumumab 184 mapatumumab 8635 Margetuximab 473 margetuximab 9799Marstacimab 852 marstacimab 10789 Maslimomab 318 maslimomab 6614Mavrilimumab 335 mavrilimumab 9234 Matuzumab 39 matuzumab 8103Mepolizumab 87 mepolizumab 7876 Metelimumab 186 metelimumab 8087Milatuzumab 135 milatuzumab 8922 Minretumomab 309 minretumomab 7821Mirikizumab 767 mirikizumab 10657 Mirvetuximab soravtansine 575mirvetuximab soravtansine 10176 Mitumomab 130 mitumomab 7934 Modotuximab433 modotuximab 9613 Mogamulizumab 366 mogamulizumab 9374 Monalizumab562 monalizumab 10113 Morolimumab 311 morolimumab 7646 Mosunetuzumab 760mosunetuzumab 10621 Motavizumab 73 motavizumab 8693 Moxetumomabpasudotox 198 moxetumomab pasudotox 9236 Muromonab-CD3 132 muromonab-CD36281 Nacolomab tafenatox 316 nacolomab tafenatox 7227 Namilumab 367namilumab 9382 Naptumomab estafenatox 65 naptumomab estafenatox 8598Naratuximab emtansine 602 naratuximab emtansine 10238 Narnatumab 381narnatumab 9447 Natalizumab 75 natalizumab 7716 Navicixizumab 597navicixizumab 10220 Navivumab 551 navivumab 9976 Nebacumab 193 nebacumab6658 Necitumumab 294 necitumumab 9083 Nemolizumab 538 nemolizumab 10064Nerelimomab 307 nerelimomab 7546 Nesvacumab 452 nesvacumab 9688Netakimab 790 netakimab 10387 Nimotuzumab 76 nimotuzumab 8545 Nirsevimab853 nirsevimab 10780 Nivolumab 424 nivolumab 9623 Obiltoxaximab 549obiltoxaximab 9825 Obinutuzumab 238 obinutuzumab 9043 Ocaratuzumab 425ocaratuzumab 9590 Ocrelizumab 227 ocrelizumab 8636 Odulimomab 134odulimomab 7364 Ofatumumab 194 ofatumumab 8606 Olaratumab 352 olaratumab9308 Oleclumab 729 oleclumab 10545 Olendalizumab 585 olendalizumab 10037Olokizumab 353 olokizumab 9333 Omalizumab 77 omalizumab 8039 Omburtamab855 omburtamab 10803 Onartuzumab 368 onartuzumab 9368 Ontuxizumab 491ontuxizumab 9519 Onvatilimab 810 onvatilimab 10758 Opicinumab 557opicinumab 10090 Oportuzumab monatox 237 oportuzumab monatox 9045Oregovomab 136 oregovomab 8183 Orticumab 426 orticumab 9635 Otelixizumab235 otelixizumab 8864 Otilimab 189 otilimab 10783 Otlertuzumab 488otlertuzumab 9832 Oxelumab 354 oxelumab 9320 Ozanezumab 454 ozanezumab9703 Ozoralizumab 382 ozoralizumab 9369 Pagibaximab 253 pagibaximab 8643Palivizumab 79 palivizumab 7753 Pamrevlumab 554 pamrevlumab 10060Panitumumab 196 panitumumab 8499 Panobacumab 243 panobacumab 8888Parsatuzumab 427 parsatuzumab 9647 Pascolizumab 88 pascolizumab 8227Pasotuxizumab 498 pasotuxizumab 9937 Pateclizumab 383 pateclizumab 9428Patritumab 407 patritumab 9549 Pembrolizumab 472 pembrolizumab 9798Perakizumab 428 perakizumab 9648 Pertuzumab 80 pertuzumab 8380Pexelizumab 81 pexelizumab 8153 Pidilizumab 453 pidilizumab 9689Pinatuzumab vedotin 457 pinatuzumab vedotin 9713 Placulumab 429placulumab 9567 Plozalizumab 566 plozalizumab 10124 Polatuzumab vedotin458 polatuzumab vedotin 9714 Ponezumab 369 ponezumab 9322 Porgaviximab777 porgaviximab 10536 Prasinezumab 769 prasinezumab 10680 Priliximab160 priliximab 7263 Pritoxaximab 459 pritoxaximab 9723 Pritumumab 270pritumumab 8132 Quilizumab 406 quilizumab 9541 Racotumomab 225racotumomab 8998 Radretumab 370 radretumab 9340 Rafivrumab 94rafivirumab 9052 Ralpancizumab 484 ralpancizumab 9841 Ramucirumab 295ramucirumab 9098 Ranevetmab 663 ranevetmab 10422 Ranibizumab 84ranibizumab 8313 Raxibacumab 260 raxibacumab 8580 Ravagalimab 806ravagalimab 10750 Ravulizumab 674 ravulizumab 10659 Refanezumab 591refanezumab 10174 Regavirumab 197 regavirumab 7250 Relatlimab 781relatlimab 10735 Remtolumab 517 remtolumab 10345 Reslizumab 279reslizumab 8106 Rilotumumab 302 rilotumumab 9123 Rinucumab 574 rinucumab10175 Risankizumab 567 risankizumab 10128 Rituximab 161 rituximab 7609Rivabazumab pegol 106 rivabazumab pegol 10144 Robatumumab 296robatumumab 9092 Roledumab 355 roledumab 9335 Romilkimab 794 romilkimab10622 Romosozumab 404 romosozumab 9533 Rontalizumab 327 rontalizumab9114 Rosmantuzumab 660 rosmantuzumab 10415 Rovalpituzumab tesirine 569rovalpituzumab tesirine 10141 Rovelizumab 304 rovelizumab 7869Rozanolixizumab 642 rozanolixizumab 10213 Ruplizumab 17 ruplizumab 8014Sacituzumab govitecan 559 sacituzumab govitecan 10097 Samalizumab 356samalizumab 9307 Samrotamab vedotin 815 samrotamab vedotin 10791Sarilumab 400 sarilumab 9476 Satralizumab 586 satralizumab 10065Secukinumab 326 secukinumab 9182 Selicrelumab 723 selicrelumab 10523Seribantumab 455 seribantumab 9710 Setoxaximab 460 setoxaximab 9724Setrusumab 632 setrusumab 10539 Sevirumab 83 sevirumab 6560 Sibrotuzumab285 sibrotuzumab 7866 Sifalimumab 322 sifalimumab 9135 Siltuximab 297siltuximab 9051 Simtuzumab 430 simtuzumab 9626 Siplizumab 71 siplizumab8251 Sirtratunab vedotin 772 sirtratumab vedotin 10467 Sirukumab 384sirukumab 9431 Sofituzumab vedotin 482 sofituzumab vedotin 9861Solanezumab 298 solanezumab 9097 Solitomab 405 solitomab 9537 Sontuzumab251 sontuzumab 8438 Spartalizumab 761 spartalizumab 10624 Stamulumab 192stamulumab 8683 Sulesomab 139 sulesomab 7519 Suptavumab 621 suptavumab10303 Sutimlimab 802 sutimlimab 10737 Suvizumab 330 suvizumab 9185Suvratoxumab 716 suvratoxumab 10441 Tabalumab 385 tabalumab 9430Tadocizumab 103 tadocizumab 8651 Talacotuzumab 754 talacotuzumab 10508Talizumab 271 talizumab 8370 Tamtuvetmab 594 tamtuvetmab 10195 Tanezumab230 tanezumab 8941 Taplitumomab paptox 282 taplitumomab paptox 8047Tarextumab 467 tarextumab 9762 Tavolimab 662 tavolimab 10420 Tefibazumab261 tefibazumab 8500 Telimomab aritox 320 telimomab aritox 6345Telisotuzumab vedotin 653 telisotuzumab vedotin 10365 Tenatumomab 226tenatumomab 8832 Teneliximab 276 teneliximab 8211 Teplizumab 92teplizumab 8869 Tepoditamab 812 tepoditamab 10766 Teprotumumab 336teprotumumab 9107 Tesidolumab 535 tesidolumab 10051 Tezepelumab 573tezepelumab 10172 Tibulizumab 776 tibulizumab 10656 Tildrakizumab 450tildrakizumab 9672 Tigatuzumab 234 tigatuzumab 8979 Timigutuzumab 739timigutuzumab 10335 Timolumab 606 timolumab 10248 Tislelizumab 757tislelizumab 10553 Tisotumab vedotin 571 tisotumab vedotin 10148Tocilizumab 96 tocilizumab 8394 Tomuzotuximab 738 tomuzotuximab 10334Toralizumab 60 toralizumab 8232 Tosatoxumab 465 tosatoxumab 9757Tositumomab 142 tositumomab 7827 Tovetumab 469 tovetumab 9778Tralokinumab 171 tralokinumab 9235 Trastuzumab 97 trastuzumab 7637Trastuzumab emtansine 357 trastuzumab emtansine 9295 Tregalizumab 371tregalizumab 9413 Tremelimumab 248 tremelimumab 8716 Trevogrumab 556trevogrumab 10087 Tucotuzumab celmoleukin 254 tucotuzumab celmoleukin8652 Tuvirumab 169 tuvirumab 6559 Ublituximab 372 ublituximab 9334Ulocuplumab 483 ulocuplumab 9854 Urelumab 373 urelumab 9365 Urtoxazumab265 urtoxazumab 8276 Ustekinumab 172 ustekinumab 8954 Utomilumab 657utomilumab 10385 Vadastuximab talirine 552 vadastuximab talirine 9983Vandortuzumab vedotin 531 vandortuzumab vedotin 10018 Vantictumab 470vantictumab 9779 Vanucizumab 500 vanucizumab 9950 Vapaliximab 277vapaliximab 8207 Varisacumab 714 varisacumab 10427 Varlilumab 497varlilumab 9933 Vatelizumab 386 vatelizumab 9439 Vedolizumab 300vedolizumab 9093 Veltuzumab 21 veltuzumab 8932 Vepalimomab 310vepalimomab 7757 Vesencumab 374 vesencumab 9380 Visilizumab 100visilizumab 8054 Vobarilizumab 523 vobarilizumab 10210 Volociximab 256volociximab 8600 Vonlerolizumab 608 vonlerolizumab 10272 Vopratelimab801 vopratelimab 10730 Vorsetuzumab mafodotin 432 vorsetuzumab mafodotin9610 Votumumab 199 votumumab 7165 Vunakizumab 658 vunakizumab 10399Xentuzumab 588 xentuzumab 10129 Zalutumumab 200 zalutumumab 8605Zanolimumab 201 zanolimumab 8298 Zenocutuzumab 771 zenocutuzumab 10687Ziralimumab 8 ziralimumab 8061 Zolbetuximab 753 zolbetuximab 10473Zolimomab aritox 314 zolimomab aritox 7055

In some embodiments, the antibody may target or bind an antigenassociated with a disease, disorder, or condition. Further, the antibodyor antibody fragment may be effective in the treatment of the disease,disorder, or condition by binding the target antigen. In someembodiments, the target of the antibody may be used to identify ordescribe said antibody. For example, 3F8 targets GD2 ganglioside fortreatment of neuroblastoma. For example, 8H19 targets B7-H13 fortreatment of neuroblastoma, sarcoma, metastatic brain cancers. Forexample, Abagovomab targets CA-125 (imitation) for treatment of ovariancancer. For example, Abciximab targets CD41 (integrin alpha-IIb) fortreatment of platelet aggregation inhibitor. For example, Abituzunmabtargets CD51 for treatment of cancer. For example, Abrezekinmab targetsinterleukin 13. For example, Abrilumab targets integrin α4β7 fortreatment of inflammatory bowel disease, ulcerative colitis, Crohn'sdisease. For example, Actoxumiab targets Clostridium difficile fortreatment of Clostridium difficile colitis. For example, Adalimumabtargets TNF-α for treatment of Rheumatoid arthritis, Crohn's disease,Plaque Psoriasis, Psoriatic Arthritis, Ankylosing Spondylitis, JuvenileIdiopathic Arthritis, Hemolytic disease of the newborn. For example,Adecatumumab targets EpCAM for treatment of prostate and breast cancer.For example, Atidortoxumab targets Staphylococcus aureusalpha toxin. Forexample, Aducanumab targets beta-amyloid for treatment of Alzheimer'sdisease. For example. Afasevikumab targets IL17A and IL17F for treatmentof multiple sclerosis. For example, Afelimomab targets TNF-α fortreatment of sepsis. For example, Alacizumab pego targets VEGFR2 fortreatment of cancer. For example, Alemtuzumab targets CD52 for treatmentof Multiple sclerosis. For example, Alirocumab targets PCSK9 fortreatment of hypercholesterolemia. For example, Altumomab pentetatetargets CEA for treatment of colorectal cancer (diagnosis). For example,Amatuximab targets mesothelin for treatment of cancer. For example,Anatumomab mafenatox targets TAG-72 for treatment of non-small cell lungcarcinoma. For example, Andecaliximab targets gelatinase B for treatmentof gastric cancer or gastroesophageal junction adenocarcinoma. Forexample, Anetumab ravtansine targets MSLN for treatment of cancer. Forexample, Anifrolumab targets interferon α/β receptor for treatment ofsystemic lupus erythematosus. For example, Anrukinzumab targets IL-13for treatment of asthma. For example, Apolizumab targets HLA-DR fortreatment of hematological cancers. For example, Aprutumab ixadotintargets FGFR2. For example, Arcitumomab targets CEA for treatment ofgastrointestinal cancers (diagnosis). For example, Ascrinvacumab targetsactivin receptor-like kinase 1 for treatment of cancer. For example,Aselizumab targets L-selectin (CD62L) for treatment of severely injuredpatients. For example, Atezolizumab targets PD-L1 for treatment ofcancer. For example, Atinumab targets RTN4. For example, Atorolimumabtargets Rhesus factor for treatment of hemolytic disease of thenewborn[citation needed]. For example, Avelumab targets PD-L1 fortreatment of cancer. For example, Azintuxizumab vedotin targets CD319for treatment of cancer. For example, Bapineuzumab targets beta amyloidfor treatment of Alzheimer's disease. For example, Basiliximab targetsCD25 (α chain of IL-2receptor) for treatment of prevention of organtransplant rejections. For example, Bavituximab targetsphosphatidylserine for treatment of cancer, viral infections. Forexample, BCD-100 targets PD-1 for treatment of melanoma. For example,Bectumomab targets CD22 for treatment of non-Hodgkin's lymphoma(detection). For example, Begelomab targets DPP4. For example,Belantamab mafodotin targets BCMA for treatment of cancer. For example,Belimumab targets BAFF for treatment of non-Hodgkin lymphoma. Forexample, Bemarituzumab targets FGFR2 for treatment of gastric cancer orgastroesophageal junction adenocarcinoma. For example, Benralizumabtargets CD125 for treatment of asthma. For example, Berlimatoxumabtargets Staphylococcus aureus bi-component leukocidin. For example,Bermekimab targets IL1A for treatment of colorectal cancer. For example,Bersanlimab targets ICAM-1. For example, Bertilimumab targets CCL11(eotaxin-1) for treatment of severe allergic disorders. For example,Besilesomab targets CEA-related antigen for treatment of inflammatorylesions and metastases (detection). For example, Bevacizumab targetsVEGF-A for treatment of metastatic cancer, retinopathy of prematurity.For example, Bezlotoxumab targets Clostridium difficile for treatment ofClostridium difficile colitis. For example, Biciromab targets fibrin II,beta chain for treatment of thromboembolism (diagnosis). For example,Bimagrumab targets ACVR2B for treatment of myostatin inhibitor. Forexample, Bimekizumab targets IL 17A and IL 17F for treatment ofankylosing spondylitis, psoriasis. For example, Birtamimab targets serumamyloid A protein for treatment of amyloidosis. For example, Bivatuzumabmertansine targets CD44 v6 for treatment of squamous cell carcinoma. Forexample, Bleselumab targets CD40 for treatment of organ transplantrejection. For example, Blinatumomab targets CD19 for treatment of pre-BALL (CD19+). For example, Blontuvetmab targets CD20. For example,Blosozumab targets SOST for treatment of osteoporosis. For example,Bococizumab targets neural apoptosis-regulated proteinase 1 fortreatment of dyslipidemia. For example, Brazikumab targets IL23 fortreatment of Crohn's disease. For example, Brentuximab vedotin targetsCD30 (TNFRSF8) for treatment of Hodgkin lymphoma, Anaplastic large-celllymphoma. For example, Briakinumab targets IL-12, IL-23 for treatment ofpsoriasis, rheumatoid arthritis, inflammatory bowel diseases, multiplesclerosis. For example, Brodalumab targets IL-17 for treatment of Plaquepsoriasis. For example, Brolucizumab targets VEGFA for treatment of wetage-related macular degeneration. For example, Brontictuzumab targetsNotch 1 for treatment of cancer. For example, Burosumab targets FGF 23for treatment of X-linked hypophosphatemia. For example, Cabiralizumabtargets CSF1R for treatment of metastatic pancreatic cancer. Forexample, Camidanlumab tesirine targets CD25 for treatment of B-cellHodgkin's lymphoma, non-Hodgkin lymphoma, acute lymphoblastic leukemia,acute myeloid leukemia. For example, Camrelizumab targets PD-1 fortreatment of hepatocellular carcinoma. For example, Canakinumab targetsIL-1 for treatment of Cryopyrin-associated periodic syndrome. Forexample, Cantuzumab mertansine targets mucin CanAg for treatment ofcolorectal cancer. For example, Cantuzumab ravtansine targets MUC1 fortreatment of cancers. For example, Caplacizumab targets VWF fortreatment of thrombotic thrombocytopenic purpura, thrombosis. Forexample, Capromab pendetide targets prostatic carcinoma cells fortreatment of prostate cancer (detection). For example, Carlumab targetsMCP-1 for treatment of oncology/immune indications. For example,Carotuximab targets endoglin for treatment of angiosarcoma. For example,Catumaxomab targets EpCAM, CD3 for treatment of ovarian cancer,malignant ascites, gastric cancer. For example, cBR96-doxorubicinimmunoconjugate targets Lewis-Y antigen for treatment of cancer. Forexample, Cedelizumab targets CD4 for treatment of prevention of organtransplant rejections, treatment of autoimmune diseases. For example,Cemiplimab targets PCDC1 for treatment of cutaneous squamous cellcarcinoma. For example, Cergutuzumab amunaleukin targets IL2 fortreatment of cancer. For example, Certolizumab pegol targets TNF-α fortreatment of Crohn's disease, Rheumatoid arthritis, axialspondyloarthritis, psoriasis arthritis. For example, Cetrelimab targetsPD-1 for treatment of cancer. For example, Cetuximab targets EGFR fortreatment of metastatic colorectal cancer and head and neck cancer. Forexample, Cibisatamab targets CEACAM5 for treatment of cancer. Forexample, Cirmtuzumab targets ROR1 for treatment of chronic lymphocyticleukemia. For example, Citatuzumab bogatox targets EpCAM for treatmentof ovarian cancer and other solid tumors. For example, Cixutumumabtargets IGF-1 receptor (CD221) for treatment of solid tumors. Forexample, Clazakizumab targets IL6 for treatment of rheumatoid arthritis.For example, Clenoliximab targets CD4 for treatment of rheumatoidarthritis. For example, Clivatuzumab tetraxetan targets MUC1 fortreatment of pancreatic cancer. For example, Codrituzumab targetsglypican 3 for treatment of cancer. For example, Cofetuzumab pelidotintargets PTK7 for treatment of cancer. For example, Coltuximab ravtansinetargets CD19 for treatment of cancer. For example, Conatumumab targetsTRAIL-R2 for treatment of cancer. For example, Concizumab targets TFPIfor treatment of bleeding. For example, Cosfroviximab targets ebolavirusglycoprotein for treatment of Ebola virus. For example, Crenezumabtargets 1-40-β-amyloid for treatment of Alzheimer's disease. Forexample, Crizanlizumab targets selectin P for treatment of sickle-celldisease. For example, Crotedumab targets GCGR for treatment of diabetes.For example, CR6261 targets Influenza A hemagglutinin for treatment ofinfectious disease/influenza A. For example, Cusatuzumab targets CD70for treatment of cancer. For example, Dacetuzumab targets CD40 fortreatment of hematologic cancers. For example, Daclizumab targets CD25(α chain of IL-2receptor) for treatment of prevention of organtransplant rejections, multiple sclerosis. For example, Dalotuzumabtargets IGF-1 receptor (CD221) for treatment of cancer. For example,Dapirolizumab pegol targets CD154 (CD40L). For example, Daratumumabtargets CD38 for treatment of Multiple myeloma. For example, Dectrekumabtargets IL-13. For example, Demcizumab targets DLL4 for treatment ofcancer. For example, Denintuzumab mafodotin targets CD19 for treatmentof cancer. For example, Denosumab targets RANKL for treatment ofosteoporosis and bone metastases. For example, Depatuxizumab mafodotintargets EGFR for treatment of glioblastoma. For example, Derlotuximabbiotin targets histone complex for treatment of recurrent glioblastomamultiforme. For example, Detumomab targets B-lymphoma cell for treatmentof lymphoma. For example, Dezamizumab targets serum amyloid P component.For example, Dinutuximab targets GD2 ganglioside for treatment ofneuroblastoma. For example, Diridavumab targets hemagglutinin fortreatment of influenza A. For example, Domagrozumab targets GDF-8 fortreatment of Duchenne muscular dystrophy. For example, Dostarlimabtargets PCDP1 for treatment of cancer. For example, Drozitumab targetsDR5 for treatment of cancer. For example, DS-8201 targets HER2 fortreatment of gastric or gastroesophageal junction adenocarcinoma. Forexample, Duligotuzumab targets ERBB3 (HER3) for treatment of testicularcancer. For example, Dupilumab targets IL4 for treatment of atopicdiseases. For example, Durvalumab targets PD-L1 for treatment of cancer.For example, Dusigitumab targets ILGF2 for treatment of B-cellmalignancies. For example, Duvortuxizumab targets CD19, CD3E fortreatment of cancer. For example, Ecromeximab targets GD3 gangliosidefor treatment of malignant melanoma. For example, Eculizumab targets C5for treatment of paroxysmal nocturnal hemoglobinuria, atypical HUS. Forexample, Edobacomab targets endotoxin for treatment of sepsis caused byGram-negative bacteria. For example, Edrecolomab targets EpCAM fortreatment of colorectal carcinoma. For example, Efalizumab targets LFA-1(CD11a) for treatment of psoriasis (blocks T-cell migration). Forexample, Efungumab targets Hsp90 for treatment of invasive Candidainfection. For example, Eldelumab targets interferon gamma-inducedprotein for treatment of Crohn's disease, ulcerative colitis. Forexample, Elezanumab targets RGMA for treatment of spinal cord injury andmultiple sclerosis. For example, Elgemtumab targets ERBB3 (HER3) fortreatment of cancer. For example, Elotuzumab targets SLAMF7 fortreatment of multiple myeloma. For example, Elsilimomab targets IL-6.For example, Emactuzumab targets CSF1R for treatment of cancer. Forexample, Emapalumab targets interferon gamma for treatment ofhemophagocytic lymphohistiocytosis. For example, Emibetuzumab targetsHHGFR for treatment of cancer. For example, Emicizumab targets activatedF9, F10 for treatment of haemophilia A. For example, Enapotamab vedotintargets AXL for treatment of cancer. For example, Enavatuzumab targetsTWEAK receptor for treatment of cancer etc. For example, Enfortumabvedotin targets nectin-4 for treatment of urothelial cancer. Forexample, Enlimomab pegol targets ICAM-1 (CD54). For example,Enoblituzumab targets CD276 for treatment of cancer. For example,Enokizumab targets IL9 for treatment of asthma. For example, Enoticumabtargets DLL4. For example, Ensituximab targets 5AC for treatment ofcancer. For example, Epitumomab cituxetan targets episialin. Forexample, Epratuzumab targets CD22 for treatment of cancer, SLE. Forexample, Eptinezumab targets calcitonin gene-related peptide fortreatment of migraine. For example, Erenumab targets CGRP for treatmentof migraine. For example, Erlizumab targets ITGB2 (CD18) for treatmentof heart attack, stroke, traumatic shock. For example, Ertumaxomabtargets HER2/neu, CD3 for treatment of breast cancer etc. For example,Etaracizumab targets integrin αvβ3 for treatment of melanoma, prostatecancer, ovarian cancer etc. For example, Etigilimab targets TIGIT. Forexample, Etrolizumab targets integrin β7 for treatment of inflammatorybowel disease. For example, Evinacumab targets angiopoietin 3 fortreatment of dyslipidemia. For example, Evolocumab targets PCSK9 fortreatment of hypercholesterolemia. For example, Exbivirumab targetshepatitis B surface antigen for treatment of hepatitis B. For example,Fanolesomab targets CD15 for treatment of appendicitis (diagnosis). Forexample, Faralimomab targets interferon receptor. For example, Faricimabtargets VEGF-A and Ang-2 for treatment of angiogenesis, ocular vasculardiseases. For example, Farletuzumab targets folate receptor 1 fortreatment of ovarian cancer. For example, Fasinumab targets HNGF fortreatment of acute sciatic pain. For example, FBTA05 targets CD20 fortreatment of chronic lymphocytic leukaemia. For example, Felvizumabtargets respiratory syncytial virus for treatment of respiratorysyncytial virus infection. For example, Fezakinumab targets IL-22 fortreatment of rheumatoid arthritis, psoriasis. For example, Fibatuzumabtargets ephrin receptor A3. For example, Ficlatuzumab targets HGF fortreatment of cancer. For example, Figitumumab targets IGF-1 receptor(CD221) for treatment of adrenocortical carcinoma and non-small celllung carcinoma. For example, Firivumab targets influenza A virushemagglutinin. For example, Flanvotumab targets TYRP1 (glycoprotein 75)for treatment of melanoma. For example, Fletikumab targets IL 20 fortreatment of rheumatoid arthritis. For example, Flotetuzumab targets IL3 receptor for treatment of hematological malignancies. For example,Fontolizumab targets IFN-γ for treatment of Crohn's disease. Forexample, Foralumab targets CD3 epsilon. For example, Foravirumab targetsrabies virus glycoprotein for treatment of rabies (prophylaxis). Forexample, Fremanezumab targets calcitonin gene-related peptide alpha fortreatment of migraine and cluster headache. For example, Fresolimumabtargets TGF-β for treatment of idiopathic pulmonary fibrosis, focalsegmental glomerulosclerosis, cancer. For example, Frovocimab targetsPCSK9 for treatment of hypercholesterolemia. For example, Frunevetmabtargets NGF. For example, Fulranumab targets NGF for treatment of pain.For example, Futuximab targets EGFR for treatment of cancer. Forexample, Galcanezumab targets calcitonin for treatment of migraine. Forexample, Galiximab targets CD80 for treatment of B-cell lymphoma. Forexample, Gancotamab targets HER2/neu for treatment of cancer. Forexample, Ganitumab targets IGF-1 receptor (CD221) for treatment ofcancer. For example, Gantenerumab targets beta amyloid for treatment ofAlzheimer's disease. For example, Gatipotuzumab targets MUC1 fortreatment of cancer. For example, Gavilimomab targets CD147 (basigin)for treatment of graft versus host disease. For example, Gedivumabtargets hemagglutinin HA. For example, Gemtuzumab ozogamicin targetsCD33 for treatment of acute myelogenous leukemia. For example,Gevokizumab targets IL-1β for treatment of diabetes. For example,Gilvetmab targets PCDC1. For example, Gimsilumab targets CSF2 fortreatment of rheumatoid arthritis. For example, Girentuximab targetscarbonic anhydrase 9 (CA-IX) for treatment of clear cell renal cellcarcinoma. For example, Glembatumumab vedotin targets GPNMB fortreatment of melanoma, breast cancer. For example, Golimumab targetsTNF-α for treatment of rheumatoid arthritis, psoriatic arthritis,ankylosing spondylitis. For example, Gomiliximab targets CD23 (IgEreceptor) for treatment of allergic asthma. For example, Gosuranemabtargets tau protein for treatment of progressive supranuclear palsy. Forexample, Guselkumab targets IL23 for treatment of psoriasis. Forexample, Ianalumab targets BAFF-R for treatment of autoimmune hepatitis.For example, Ibalizumab targets CD4 for treatment of HIV infection. Forexample, IBI308 targets PD-1 for treatment of squamous cell non-smallcell lung cancer. For example, Ibritumomab tiuxetan targets CD20 fortreatment of non-Hodgkin's lymphoma. For example, Icrucumab targetsVEGFR-1 for treatment of cancer. For example, Idarucizumab targetsdabigatran for treatment of reversal of anticoagulant effects ofdabigatran. For example, Ifabotuzumab targets EPHA3. For example,Igovomab targets CA-125 for treatment of ovarian cancer (diagnosis). Forexample, Iladatuzumab vedotin targets CD97B for treatment of cancer. Forexample, IMAB362 targets CLDN18.2 for treatment of gastrointestinaladenocarcinomas and pancreatic tumor. For example, Imalumab targets MIFfor treatment of cancer. For example, Imaprelimab targets MCAM. Forexample, Imciromab targets cardiac myosin for treatment of cardiacimaging. For example, Imgatuzumab targets EGFR for treatment of cancer.For example, Inclacumab targets selectin P for treatment ofcardiovascular disease. For example, Indatuximab ravtansine targets SDC1for treatment of cancer. For example, Indusatumab vedotin targets GUCY2Cfor treatment of cancer. For example, Inebilizumab targets CD19 fortreatment of cancer, systemic sclerosis, multiple sclerosis. Forexample, Infliximab targets TNF-α for treatment of rheumatoid arthritis,ankylosing spondylitis, psoriatic arthritis, psoriasis, Crohn's disease,ulcerative colitis. For example, Intetumumab targets CD51 for treatmentof solid tumors (prostate cancer, melanoma). For example, Inolimomabtargets CD25 (α chain of IL-2receptor) for treatment of graft versushost disease. For example, inotuzumab ozogamicin targets CD22 fortreatment of ALL. For example, Ipilimumab targets CD152 for treatment ofmelanoma. For example, Iomab-B targets CD45 for treatment of ablation ofbone marrow. For example, Iratumumab targets CD30 (TNFRSF8) fortreatment of Hodgkin's lymphoma. For example, Isatuximab targets CD38for treatment of multiple myeloma. For example, Iscalimab targets CD40.For example, Istiratumab targets IGF1R, CD221 for treatment of advancedsolid tumors. For example, Itolizumab targets CD6 for treatment ofpsoriasis. For example, Ixekizumab targets IL 17A for treatment ofautoimmune diseases. For example, Keliximab targets CD4 for treatment ofchronic asthma. For example, Labetuzumab targets CEA for treatment ofcolorectal cancer. For example, Lacnotuzumab targets CSF1, MCSF fortreatment of cancer. For example, Ladiratuzumab vedotin targets LIV-1for treatment of cancer. For example, Lampalizumab targets CFD fortreatment of geographic atrophy secondary to age-related maculardegeneration. For example, Lanadelumab targets kallikrein for treatmentof angioedema. For example, Landogrozumab targets GDF-8 for treatment ofmuscle wasting disorders. For example, Laprituximab emtansine targetsEGFR For example, Larcaviximab targets ebolavirus glycoprotein fortreatment of Ebola virus. For example, Lebrikizumab targets IL-13 fortreatment of asthma. For example, Lemalesomab targets NCA-90(granulocyte antigen) for treatment of diagnostic agent. For example,Lendalizumab targets C5. For example, Lenvervimab targets hepatitis Bsurface antigen for treatment of hepatitis B. For example, Lenzilumabtargets CSF2 for treatment of chronic myelomonocytic leukemia andjuvenile myelomonocytic leukemia. For example, Lerdelimumab targets TGFbeta 2 for treatment of reduction of scarring after glaucoma surgery.For example, Leronlimab targets CCR5. For example, Lesofavumab targetshemagglutinin HA. For example, Letolizumab targets TRAP for treatment ofinflammatory diseases. For example, Lexatumumab targets TRAIL-R2 fortreatment of cancer. For example, Libivirumab targets hepatitis Bsurface antigen for treatment of hepatitis B. For example, Lifastuzumabvedotin targets phosphate-sodium co-transporter for treatment of cancer.For example, Ligelizumab targets IGHE for treatment of severe asthma andchronic spontaneous urticaria. For example, Loncastuximab tesirinetargets CD19 for treatment of cancer. For example, Losatuxizumab vedotintargets EGRF, ERBB1 HER1 for treatment of cancer. For example, Lilotomabsatetraxetan targets CD37 for treatment of cancer. For example,Lintuzumab targets CD33 for treatment of cancer. For example, Lirilumabtargets KIR2D for treatment of solid and hematological cancers. Forexample, Lodelcizumab targets PCSK9 for treatment ofhypercholesterolemia. For example, Lokivetmab targets Canis lupusfamiliaris IL31 for treatment of clinical signs of atopic dermatitis indogs[50]. For example, Lorvotuzumab mertansine targets CD56 fortreatment of cancer. For example, Lucatumumab targets CD40 for treatmentof multiple myeloma, non-Hodgkin's lymphoma, Hodgkin's lymphoma. Forexample, Lulizumab pegol targets CD28 for treatment of autoimmunediseases. For example, Lumiliximab targets CD23 (IgE receptor) fortreatment of chronic lymphocytic leukemia. For example, Lumretuzumabtargets ERBB3 (HER3) for treatment of cancer. For example, Lupartumabamadotin targets LYPD3. For example, Lutikizumab targets interleukin 1alpha. For example, Mapatumumab targets TRAIL-R1 for treatment ofcancer. For example, Margetuximab targets HER2 for treatment of breastcancer. For example, Marstacimab targets TFPI for treatment of bleedingwith hemophilia. For example, Maslimomab targets T-cell receptor. Forexample, Mavrilimumab targets GMCSF receptor α-chain for treatment ofrheumatoid arthritis. For example, Matuzumab targets EGFR for treatmentof colorectal, lung and stomach cancer. For example, Mepolizumab targetsIL-5 for treatment of asthma and white blood cell diseases. For example,Metelimumab targets TGF beta 1 for treatment of systemic scleroderma.For example, Milatuzumab targets CD74 for treatment of multiple myelomaand other hematological malignancies. For example, Minretumomab targetsTAG-72 for tumor detection and therapy. For example, Mirikizumab targetsIL23A for treatment of psoriasis. For example, Mirvetuximab soravtansinetargets folate receptor alpha for treatment of ovarian cancer. Forexample, Mitumomab targets GD3 ganglioside for treatment of small celllung carcinoma. For example, Modotuximab targets EGFR extracellulardomain III for treatment of cancer. For example, Mogamulizumab targetsCCR4 for treatment of adult T-cell leukemia/lymphoma. For example,Monalizumab targets NKG2A for treatment of rheumatoid arthritis,gynecologic malignancies, and other cancers. For example, Morolimumabtargets Rhesus factor. For example, Mosunetuzumab targets CD3E, MS4A1,CD20 for treatment of cancer. For example, Motavizumab targetsrespiratory syncytial virus for treatment of respiratory syncytial virus(prevention). For example, Moxetumomab pasudotox targets CD22 fortreatment of hairy cell leukemia. For example, Muromonab-CD3 targets CD3for treatment of prevention of organ transplant rejections. For example,Nacolomab tafenatox targets C242 antigen for treatment of colorectalcancer. For example, Namilumab targets CSF2. For example, Naptumomabestafenatox targets 5T4 for treatment of non-small cell lung carcinoma,renal cell carcinoma. For example, Naratuximab emtansine targets CD37for treatment of. For example, Narnatumab targets MST1R (aka RON) fortreatment of cancer. For example, Natalizumab targets integrin α4 fortreatment of multiple sclerosis, Crohn's disease. For example,Navicixizumab targets DLL4 and VEGFA for treatment of cancer. Forexample, Navivumab targets influenza A virushemagglutinin HA. Forexample, Naxitamab targets c-Met for treatment of high-riskneuroblastoma and refractory osteomedullary disease. For example,Nebacumab targets endotoxin for treatment of sepsis. For example,Necitumumab targets EGFR for treatment of non-small cell lung carcinoma.For example, Nemolizumab targets IL31RA for treatment of eczema[53]. Forexample, NEOD001 targets amyloid for treatment of primary systemicamyloidosis. For example, Nerelimomab targets TNF-α. For example,Nesvacumab targets angiopoietin 2 for treatment of cancer. For example,Netakimab targets interleukin 17A. For example, Nimotuzumab targets EGFRfor treatment of squamous cell carcinoma, head and neck cancer,nasopharyngeal cancer, glioma. For example, Nirsevimab targets RSVFR fortreatment of respiratory syncytial virus. For example, Nivolumab targetsPD-1 for treatment of cancer. For example, Nofetumomab merpentan treatscancer (diagnosis). For example, Obiltoxaximab targets Bacillusanthracis anthrax for treatment of Bacillus anthracis spores. Forexample, Obinutuzumab targets CD20 for treatment of Chronic lymphaticleukemia. For example, Ocaratuzumab targets CD20 for treatment ofcancer. For example, Ocrelizumab targets CD20 for treatment ofrheumatoid arthritis, lupus erythematosus. For example, Odulimomabtargets LFA-1 (CD11a) for treatment of prevention of organ transplantrejections, immunological diseases. For example, Ofatumumab targets CD20for treatment of chronic lymphocytic leukemia. For example, Olaratumabtargets PDGF-R α for treatment of cancer. For example, Oleclumab targets5′-nucleotidase for treatment of pancreatic and colorectal cancer. Forexample, Olendalizumab targets complement C5a for treatment of systemiclupus erythematosus, lupus nephritis, acute graft-versus-host disease.For example, Olokizumab targets IL6 for treatment of rheumatoidarthritis. For example, Omalizumab targets IgE Fc region for treatmentof allergic asthma. For example, Omburtamab targets CD276 for treatmentof cancer. For example, OMS721 targets MASP-2 for treatment of atypicalhemolytic uremic syndrome. For example, Onartuzumab targets humanscatter factor receptor kinase for treatment of cancer. For example,Ontuxizumab targets TEM1 for treatment of cancer. For example,Onvatilimab targets VSIR For example, Opicinumab targets LINGO-1 fortreatment of multiple sclerosis. For example, Oportuzumab monatoxtargets EpCAM for treatment of bladder cancer. For example, Oregovomabtargets CA-125 for treatment of ovarian cancer. For example, Orticumabtargets oxLDL. For example, Otelixizumab targets CD3 for treatment ofdiabetes mellitus type 1. For example, Otilimab targets GMCSF fortreatment of osteoarthritis, rheumatoid arthritis. For example,Otlertuzumab targets CD37 for treatment of cancer. For example, Oxelumabtargets OX-40 for treatment of asthma. For example, Ozanezumab targetsNOGO-A for treatment of ALS and multiple sclerosis. For example,Ozoralizumab targets TNF-α for treatment of inflammation. For example,Pagibaximab targets lipoteichoic acid for treatment of sepsis(Staphylococcus). For example, Palivizumab targets F protein ofrespiratory syncytial virus for treatment of respiratory syncytial virus(prevention). For example, Pamrevlumab targets CTGF for treatment ofidiopathic pulmonary fibrosis (IPF), pancreatic cancer. For example,Panitumumab targets EGFR for treatment of colorectal cancer. Forexample, Pankomab targets tumor specific glycosylation of MUC1 fortreatment of ovarian cancer. For example, Panobacumab targetsPseudomonas aeruginosa for treatment of Pseudomonas aeruginosainfection. For example, Parsatuzumab targets EGFL7 for treatment ofcancer. For example, Pascolizumab targets IL-4 for treatment of asthma.For example, Pasotuxizumab targets folate hydrolase for treatment ofcancer. For example, Pateclizumab targets LTA for treatment of TNF. Forexample, Patritumab targets ERBB3 (HER3) for treatment of cancer. Forexample, PDR001 targets PD-1 for treatment of melanoma. For example,Pembrolizumab targets PD-1 for treatment of melanoma and other cancers.For example, Pemtumomab targets MUC1 for treatment of cancer. Forexample, Perakizumab targets IL 17A for treatment of arthritis. Forexample, Pertuzumab targets HER2/neu for treatment of cancer. Forexample, Pexelizumab targets C5 for treatment of reduction of sideeffects of cardiac surgery. For example, Pidilizumab targets PD-1 fortreatment of cancer and infectious diseases. For example, Pinatuzumabvedotin targets CD22 for treatment of cancer. For example, Pintumomabtargets adenocarcinoma antigen for treatment of adenocarcinoma(imaging). For example, Placulumab targets human TNF for treatment ofpain and inflammatory diseases. For example, Plozalizumab targets CCR2for treatment of diabetic nephropathy and arteriovenous graft patency.For example, Pogalizumab targets TNFR superfamily member 4. For example,Polatuzumab vedotin targets CD79B for treatment of diffuse large B-celllymphoma. For example, Ponezumab targets human beta-amyloid fortreatment of Alzheimer's disease. For example, Porgaviximab targetsZaire ebolavirus glycoprotein for treatment of Ebola virus disease. Forexample, Prasinezumab targets NACP for treatment of Parkinson's disease.For example, Prezalizumab targets ICOSL. For example, Priliximab targetsCD4 for treatment of Crohn's disease, multiple sclerosis. For example,Pritoxaximab targets E. coli shiga toxin type-1. For example, Pritumumabtargets vimentin for treatment of brain cancer. For example, PRO 140targets CCR5 for treatment of HIV infection. For example, Quilizumabtargets IGHE for treatment of asthma. For example, Racotumomab targetsNGNA ganglioside for treatment of non-small cell lung cancer. Forexample, Radretumab targets fibronectin extra domain-B for treatment ofcancer. For example, Rafivirumab targets rabies virus glycoprotein fortreatment of rabies (prophylaxis). For example, Ralpancizumab targetsneural apoptosis-regulated proteinase 1 for treatment of dyslipidemia.For example, Ramucirumab targets VEGFR2 for treatment of solid tumors.For example, Ranevetmab targets NGF for treatment of osteoarthritis indogs. For example, Ranibizumab targets VEGF-A for treatment of maculardegeneration (wet form). For example, Raxibacumab targets anthrax toxin,protective antigen for treatment of anthrax (prophylaxis and treatment).For example, Ravagalimab targets CD40 for treatment of Crohn's disease.For example, Ravulizumab targets C5 for treatment of paroxysmalnocturnal hemoglobinuria, atypical hemolytic uremic syndrome. Forexample, Refanezumab targets myelin-associated glycoprotein fortreatment of recovery of motor function after stroke. For example,Regavirumab targets cytomegalovirus glycoprotein B for treatment ofcytomegalovirus infection. For example, Relatlimab targets LAG3 fortreatment of melanoma. For example, Remtolumab targets interleukin 17alpha (TNF-α). For example, Reslizumab targets IL-5 for treatment ofinflammations of the airways, skin and gastrointestinal tract. Forexample, Rilotumumab targets HGF for treatment of solid tumors. Forexample, Rinucumab targets platelet-derived growth factor receptor betafor treatment of neovascular age-related macular degeneration. Forexample, Risankizumab targets IL23A for treatment of Crohn's disease,psoriasis, psoriatic arthritis, and asthma. For example, Rituximabtargets CD20 for treatment of lymphomas, leukemias, some autoimmunedisorders. For example, Rivabazumab pegol targets Pseudomonas aeruginosatype III secretion system. For example, Robatumumab targets IGF-1receptor (CD221) for treatment of cancer. For example, Rmab targetsrabies virus G glycoprotein for treatment of post-exposure prophylaxisof rabies. For example, Roledumab targets RHD for treatment of Rhdisease. For example, Romilkimab targets interleukin 13. For example,Romosozumab targets sclerostin for treatment of osteoporosis. Forexample, Rontalizumab targets IFN-α for treatment of systemic lupuserythematosus. For example, Rosmantuzumab targets root plate-specificspondin 3 for treatment of cancer. For example, Rovalpituzumab tesirinetargets DLL3 for treatment of small cell lung cancer. For example,Rovelizumab targets CD11, CD18 for treatment of hemorrhagic shock. Forexample, Rozanolixizumab targets FCGRT. For example, Ruplizumab targetsCD154 (CD40L) for treatment of rheumatic diseases. For example, SA237targets IL-6R for treatment of neuromyelitis optica and neuromyelitisoptica spectrum disorders. For example, Sacituzumab govitecan targetsTROP-2 for treatment of triple-negative breast cancer. For example,Samalizumab targets CD200 for treatment of cancer. For example,Samrotamab vedotin targets LRRC15 for treatment of cancer. For example,Sarilumab targets IL6 for treatment of rheumatoid arthritis, ankylosingspondylitis. For example, Satralizumab targets IL6 receptor fortreatment of neuromyelitis optica. For example, Satumomab pendetidetargets TAG-72 for treatment of cancer (diagnosis). For example,Secukinumab targets IL 17A for treatment of uveitis, rheumatoidarthritis psoriasis. For example, Selicrelumab targets CD40. Forexample, Seribantumab targets ERBB3 (HER3) for treatment of cancer. Forexample, Setoxaximab targets E. coli shiga toxin type-2. For example,Setrusumab targets SOST. For example, Sevirumab targets cytomegalovirusfor treatment of cytomegalovirus infection. For example, Sibrotuzumabtargets FAP for treatment of cancer. For example, SGN-CD19A targets CD19for treatment of acute lymphoblastic leukemia and B-cell non-Hodgkinlymphoma. For example, SHP647 targets mucosal addressin cell adhesionmolecule for treatment of Crohn's disease. For example, Sifalimumabtargets IFN-α for treatment of SLE, dermatomyositis, polymyositis. Forexample, Siltuximab targets IL-6 for treatment of cancer. For example,Simtuzumab targets LOXL2 for treatment of fibrosis. For example,Siplizumab targets CD2 for treatment of psoriasis, graft-versus-hostdisease (prevention). For example, Sirtratumab vedotin targets SLITRK6for treatment of cancer. For example, Sirukumab targets IL-6 fortreatment of rheumatoid arthritis. For example, Sofituzumab vedotintargets CA-125 for treatment of ovarian cancer. For example, Solanezumabtargets beta amyloid for treatment of Alzheimer's disease. For example,Solitomab targets EpCAM for treatment of gastrointestinal, lung, andother cancers. For example, Sonepcizumab targets sphingosine-1-phosphatefor treatment of choroidal and retinal neovascularization. For example,Sontuzumab targets episialin. For example, Spartalizumab targets PDCD1,CD279 for treatment of melanoma. For example, Stamulumab targetsmyostatin for treatment of muscular dystrophy. For example, Sulesomabtargets NCA-90 (granulocyte antigen) for treatment of osteomyelitis. Forexample, Suptavumab targets RSVFR for treatment of medically attendedlower respiratory disease. For example, Sutimlimab targets C1s fortreatment of cold agglutinin disease. For example, Suvizumab targetsHIV-1 for treatment of viral infections. For example, Suvratoxumabtargets Staphylococcus aureusalpha toxin for treatment of nosocomialpneumonia. For example, Tabalumab targets BAFF for treatment of B-cellcancers. For example, Tacatuzumab tetraxetan targets alpha-fetoproteinfor treatment of cancer. For example, Tadocizumab targets integrinαIIbβ3 for treatment of percutaneous coronary intervention. For example,Talacotuzumab targets CD123. For example, Talizumab targets IgE fortreatment of allergic reaction. For example, Tamtuvetmab targets CD52for treatment of. For example, Tanezumab targets NGF for treatment ofpain. For example, Taplitumomab paptox targets CD19 for treatment ofcancer. For example, Tarextumab targets Notch receptor for treatment ofcancer. For example, Tavolimab targets CD134 for treatment of cancer.For example, Tefibazumab targets clumping factor A for treatment ofStaphylococcus aureus infection. For example, Telisotuzumab vedotintargets HGFR for treatment of cancer. For example, Tenatumomab targetstenascin C for treatment of cancer. For example, Teneliximab targetsCD40 for treatment of autoimmune diseases and prevention of organtransplant rejection. For example, Teplizumab targets CD3 for treatmentof diabetes mellitus type 1. For example, Tepoditamab targets dendriticcell-associated lectin 2 for treatment of cancer. For example,Teprotumumab targets IGF-1 receptor (CD221) for treatment of thyroid eyedisease. For example, Tesidolumab targets C5. For example, Tetulomabtargets CD37 for treatment of cancer. For example, Tezepelumab targetsTSLP for treatment of asthma, atopic dermatitis. For example, TGN1412targets CD28 for treatment of chronic lymphocytic leukemia, rheumatoidarthritis. For example, Tibulizumab targets BAFF for treatment ofautoimmune disorders. For example, Tildrakizumab targets IL23 fortreatment of immunologically mediated inflammatory disorders. Forexample, Tigatuzumab targets TRAIL-R2 for treatment of cancer. Forexample, Timigutuzumab targets HER2 for treatment of cancer. Forexample, Timolumab targets AOC3. For example, Tiragotumab targets TIGITfor treatment of cancer. For example, Tislelizumab targets PCDC1, CD279for treatment of non-small cell lung cancer. For example, Tisotumabvedotin targets coagulation factor III for treatment of relapsed orrefractory cervical cancer for example, TNX-650 targets IL-13 fortreatment of Hodgkin's lymphoma. For example, Tocilizumab targets IL-6receptor for treatment of rheumatoid arthritis. For example,Tomuzotuximab targets EGFR, HER1 for treatment of cancer. For example,Toralizumab targets CD154 (CD40L) for treatment of rheumatoid arthritis,lupus nephritis. For example, Tosatoxumab targets Staphylococcus aureus.For example, Tositumomab targets CD20 for treatment of follicularlymphoma. For example, Tovetumab targets PDGFRA for treatment of cancer.For example, Tralokinumab targets IL-13 for treatment of asthma, atopicdermatitis. For example, Trastuzumab targets HER2/neu for treatment ofbreast cancer. For example, Trastuzumab emtansine targets HER2/neu fortreatment of breast cancer. For example, TRBS07 targets GD2 gangliosidefor treatment of melanoma. For example, Tregalizumab targets CD4 fortreatment of. For example, Tremelimumab targets CTLA-4 for treatment ofnon-small cell lung, head & neck, urothelial cancer. For example,Trevogrumab targets growth differentiation factor8 for treatment ofmuscle atrophy due to orthopedic disuse and sarcopenia. For example,Tucotuzumab celmoleukin targets EpCAM for treatment of cancer. Forexample, Tuvirumab targets hepatitis B virus for treatment of chronichepatitis B. For example, Ublituximab targets MS4A1 for treatment ofmultiple sclerosis, chronic lymphocytic leukemia. For example,Ulocuplumab targets CXCR4 (CD184) for treatment of hematologicmalignancies. For example, Urelumab targets 4-1BB (CD137) for treatmentof cancer. For example, Urtoxazumab targets Escherichia coli fortreatment of diarrhea caused by E. coli. For example, Ustekinumabtargets IL-12, IL-23 for treatment of multiple sclerosis, psoriasis,psoriatic arthritis. For example, Utomilumab targets 4-1BB (CD137) fortreatment of diffuse large B-cell lymphoma. For example, Vadastuximabtalirine targets CD33 for treatment of Acute myeloid leukemia. Forexample, Vanalimab targets CD40. For example, Vandortuzumab vedotintargets STEAP1 for treatment of cancer. For example, Vantictumab targetsFrizzled receptor for treatment of cancer. For example, Vanucizumabtargets angiopoietin 2 for treatment of cancer. For example, Vapaliximabtargets AOC3 (VAP-1). For example, Varisacumab targets VEGF-A fortreatment of angiogenesis. For example, Varlilumab targets CD27 fortreatment of solid tumors and hematologic malignancies. For example,Vatelizumab targets ITGA2 (CD49b). For example, Vedolizumab targetsintegrin α4β7 for treatment of Crohn's disease, ulcerative colitis. Forexample, Veltuzumab targets CD20 for treatment of non-Hodgkin'slymphoma. For example, Vepalimomab targets AOC3 (VAP-1) for treatment ofinflammation. For example, Vesencumab targets NRP1 for treatment ofsolid malignancies. For example, Visilizumab targets CD3 for treatmentof Crohn's disease, ulcerative colitis. For example, Vobarilizumabtargets IL6R for treatment of inflammatory autoimmune diseases. Forexample, Volociximab targets integrin α5β1 for treatment of solidtumors. For example, Vonlerolizumab targets CD134 for treatment ofcancer. For example, Vopratelimab targets CD278, aka ICOS. For example,Vorsetuzumab mafodotin targets CD70 for treatment of cancer. Forexample, Votumumab targets tumor antigen CTAA16.88 for treatment ofcolorectal tumors. For example, Vunakizumab targets interleukin 17alpha. For example, Xentuzumab targets IGF1, IGF2. For example,XMAB-5574 targets CD19 for treatment of diffuse large B-cell lymphoma.For example, Zalutumumab targets EGFR for treatment of squamous cellcarcinoma of the head and neck. For example, Zanolimumab targets CD4 fortreatment of rheumatoid arthritis, psoriasis, T-cell lymphoma. Forexample, Zatuximab targets HER1 for treatment of cancer. For example,Zenocutuzumab targets ERBB3, HER3 for treatment of cancer. For example,Ziralimumab targets CD147 (basigin). For example, Zolbetuximab targetsCLDN18 for treatment of cancer. For example, Zolimomab aritox targetsCD5 for treatment of systemic lupus erythematosus, graft-versus-hostdisease.

In some embodiments, the antibody or antibody fragment may be human.Alternatively, the antibody or the antibody fragment may be from amouse. In some embodiments, the antibody or the antibody fragment may behumanized.

In some embodiments, the antibody or antibody fragment may bind aprotein selected from Table 2. In some embodiments, the antibody or theantibody fragment may bind a protein encoded by IL2 (interleukin 2;ENSG00000109471). In some embodiments, the antibody or antibody fragmentmay bind a histone complex. In some embodiments, the antibody orantibody fragment may bind a protein encoded by kallikrein (KLK;ENSG00000167759). In some embodiments, the antibody or antibody fragmentmay bind amyloid. In some embodiments, the antibody or antibody fragmentmay bind a Notch receptor. In some embodiments, the antibody or antibodyfragment may bind a protein encoded by oxidized low density receptor 1(OLR1; ENSG00000173391).

3. Signaling Pathways

Engineered platelets described herein may contain genetic modificationswithin the gene components of pathways for platelet adhesion, migration,and extravasation, or the engineered platelets may be loaded withproteins, nucleic acids, or small molecule drugs. The engineeredplatelets may not respond to endogenous stimuli usually resulting inclot formation, may not be recruited by other activated platelets, andon activation, may not be able to recruit and activate endogenousplatelets in the patient.

Alternatively, the deletion or modification is introduced to genes thatmediate platelet signal transduction, such as HPS (biogenesis oflysosomal organelles complex 3 subunit) genes, which are vital to ADP,serotonin, and ATP release from dense granules; and mitochondriallyencoded cytochrome C oxidase II (COX2), which generates inflammatory andprothrombogenic mediators and is a target of aspirin. Alternatively, thedeletion or modification is introduced to genes expressing thromboticmediators, such as prothrombin (major protein thrombotic inducer); PDGFwhich is a pro-angiogenic factor; EGF (elongation growth factor); andvon Willebrand Factor (collagen adaptor protein).

The combinatorial loss of thrombin and ADP signaling has been observedto abrogate vessel occlusion, but ITAM receptors can still be activated(See, Boulaftali et al. “Platelet ITAM signaling is critical forvascular integrity in inflammation”. JCI, 2013 and Cornelissen et al.“Roles and interactions among protease-activated receptors and P2ry12 inhemostasis and thrombosis”, PNAS. 2010, each of which is herebyincorporated by reference in its entirety). This work demonstrates thatdisruption of crucial endogenous platelet signaling pathways does notabrogate a platelet's ability to signal through ITAM receptors,indicating that the engineered CPRs described herein are likely tofunction on a non-thrombogenic platelet background.

For example, thrombin activates platelets through cleavage of PARs(protease activated receptors). Platelet signaling is also driven byprotease activated GPCRs, namely PAR1 and PAR4 which are cleaved bythrombin. Signaling is potent and acts to recruit platelets andfacilitate positive feedback between platelets after plateletactivation. The thrombin cleavage sequence on PAR1 and PAR4 is welldefined.

In some embodiments, the engineered platelets described herein maycomprise at least one deletion or modification introduced into orreplacing domains of endogenous platelet receptors, such as, but notlimited to, PAR4 (protease activated receptor 4), which is a primarythrombin receptor; GPIb-1X-V (Glycoprotein Ib complexed withglycoprotein IX), which is a primary anchor receptor; P2Y12 (purinergicreceptor P2Y12), which is an ADP (adenosine diphosphate) receptor andtarget of clopidogrel inhibition; GPVI (glycoprotein deletiontein VIplatelet), which is a collagen receptor; or a thromboxan receptor toprevent activation of the engineered platelet.

In some embodiments, the engineered platelets can synthesize protein inresponse to an activation signal. For example, in Weyrich et al., BCL-3was specifically upregulated in activated platelets through amechanistic target of rapamycin (mTOR) dependent signaling mechanism(See, Weyrich et al. “Signal-dependent translation of a regulatoryprotein, Bcl-3, in activated human platelets”. PNAS, 1998, which ishereby incorporated by reference in its entirety). Therefore, knock-inof a gene into the BCL-3 locus or identification of the minimal 5′ UTRregion that mediates activation dependent translation would allowsynthetic gene expression regulation in platelets. Therefore, plateletsdescribed herein may have an altered signaling pathway resulting insignaling induced protein translation. For example, expressing a toxicprotein once activated or triggering downstream events following targetcell recognition.

4. Proteins Associated with Autoimmunity

In some embodiments, a CPR of the engineered platelets described hereinmay comprise at least a portion of a protein associated withautoimmunity. For example, the CPR may comprise at least a portion of aprotein selected from the group consisting of: myelin oligodendrocyteglycoprotein (MOG), glutamic acid decarboxylase 2 (GAD65), myelinassociated glycoprotein (MAG), peripheral myelin protein 22 (PMP22),thyroid peroxidase (TPO), voltage-gated potassium channel (VGKC),proteolipid protein (PLP), acetylcholine receptor (AChR), tribblespseudokinase 2 (TRIB2), N-methyl-D-aspartate (NMDA)-type glutamatereceptor (GluR), glutamate decarboxylase 2 (GAD2), Armadillo repeatcontaining 9 (ARMC9), Cytochrome P450 Family 21 Subfamily A Member 2(CYP21A2), calcium sensing receptor (CASR), nuclear autoantigenic spermprotein (NASP), insulin, thyroid stimulating hormone receptor (TSHR),thyroperoxidase, asioglycoprotein receptor, Cytochrome P450 Family 2Subfamily D Member 6 (CYP2D6), lactoferrin (LF), tissuetrans-glutaminase (TTG), H/K ATP-ase, Factor XIII (F8),beta2-glycoprotein I (Beta2-GPI), erythrocyte I/I, B2 integrin (ITGB2),granulocyte-colony stimulating factor (G-CSF), glycoprotein (GP)IIb/IIa, collagen II (COLII), fibrinogen (FBG) βα, myeloperoxidase(MPO), cardiac myosin (CYO), proteinase 3 (PRTN3), trichohyalin (TCHH),bullous pemphigoid associated (BP), glycoprotein 1 (GP1), laminin-332(LM332), transglutaminase (TGM), type VII collagen (COLVII), P80 Coilin(COIL), Desmoglein I (DSG1), Desmoglein III (DSG3), SRY-Box 10 (SOX10),small nuclear ribonucleoprotein U1 subunit (70SNRNP70), S-antigen (SAG),and Collagen alpha-3(IV) chain (α3(IV)NC1 collagen). For example,desmoglein3-ITAM CPR may be used to target pemphigus vulgaris B cells.Alternatively, the engineered platelets described herein express an MHCclass 1-ITAM chimeric platelet receptor or MHC class 2-ITAM chimericplatelet receptor, such that the MHC class 1 or the MHC class 2 may beloaded with a peptide from the list above on the surface of the plateletto target autoimmune mediating T cells for destruction or forsuppression through the release of anti-inflammatory cytokines, such asTGF-β. Additionally, RNA encoding transcription factors may be released,such as FOXP3 to transdifferentiate bound T cells into TRegs

C. Universal Platelets

In some embodiments, the engineered platelets described herein are lessimmunogenic than platelets produced in vivo, (e.g., platelets from ahuman donor). In vitro generated platelets may be made universal throughdeletion of the 32 microglobulin gene (See, Feng et al. “ScalableGeneration of Universal Platelets from Human Induced Pluripotent StemCells”. Stem Cell Reports, 2014, which is hereby incorporated byreference in its entirety). Even without this deletion, platelets withABO matching are generally used in clinical practice without adverseeffects. O-type platelets from humans are not universal donors as theyare contaminated with anti-A/B antibodies, but contamination would notbe an issue with in vitro platelets. Therefore, in certain embodiments,the inventions described herein may use these technologies to massproduce gene-edited platelets, which are also easily stored,transported, and do not require patient matching.

D. Cargo

Platelets naturally absorb drugs and antibodies in their environmentthrough endocytosis and the open canalicular system (See, Xu et al.“Doxorubicin-loaded platelets as a smart drug delivery system: Animproved therapy for lymphoma”. Scientific reports, 2017 and Verheul etal. “Platelets Take Up the Monoclonal Antibody Bevacizumab”. HumanCancer Biology, 2007, which of which is hereby incorporated by referencein its entirety). Platelets may be used to deliver passively loaded andgenetically encoded therapeutic agents. For example, the engineeredplatelets may be passively loaded with therapeutic agents throughendocytosis and absorption. In fact, platelet α-granules contain proteineffectors and loading of soluble proteins is performed through a simplesignal peptide. A minimal targeting sequence for directing proteins intoplatelet secretory α-granules has been previously defined (See, Golli etal. “Evidence for a Granule Targeting Sequence within Platelet Factor4.”, JBC, 2004, which is hereby incorporated by reference in itsentirety). In some embodiments, activation trigger drug release in theengineered platelets. Cargo may be soluble or membrane-bound.

The cargo may also be an imaging agent.

In some preferred embodiments the cargo is not an agent that isnaturally found within the platelet, i.e. the cargo is an exogenouscargo rather than an endogenous cargo with respect to the platelet. Theskilled person will appreciate that a cargo can be exogenous to theplatelet but endogenous to the subject.In some preferred embodiments the cargo is not an agent that isnaturally found within the platelet α-granule. For example the cargo maybe an agent that is naturally found within the platelet, but notnaturally found within the α-granule.In some embodiments the cargo may be an agent that is endogenously foundwithin the platelet but is found at a higher concentration or amountwithin the platelet, or within the α-granule of the platelet than in aplatelet not of the invention.In some embodiments the cargo comprises an α-granule localization signalwherein the α-granule localization signal directs the cargo to uptakeinto α-granule vesicles of the engineered platelet. For example in someembodiments a therapeutic agent or an imaging agent comprises or isconjugated to an α-granule localization signal.

1. Toxins

In some embodiments, engineered platelets may be loaded with toxin,which would be cloaked from the immune system. The engineered plateletsmay also be loaded with chemokines and/or selectins to mediate transferof an agent across the blood brain barrier (BBB). Other embodiments ofthe engineered platelets may have platelet secretory granules loadedwith membrane and/or soluble proteins. In certain embodiments, a toxinmay be encoded with an α-granule localization signal attached to directits uptake into secretory granules, which would be released on plateletreceptor activation.

Platelet expression of programmed cell death protein (PD-1) and loadingof an engineered platelet with cyclophosphamide has been observed tofunction as a potent anti-melanoma agent (See, Zhang et al. “EngineeringPD-1-Presenting Platelets for Cancer Immunotherapy.” Nano Letters, 2018,which is hereby incorporated by reference in its entirety).Specifically, megakaryocytes were engineered to express PD-1, then theresulting engineered platelets were passively loaded withcyclophosphamide. Platelet targeting to the melanoma was driven bysurgical wounding of the tumor in vivo, not a synthetic receptor,resulting in T_(reg) depletion in the tumor and increased CD8⁺ T cellmediated killing. Tumor volume was observed to be significantly less 20days after the beginning treatment for animals in the group with bothPD-1 and cyclophosphamide compared to animals treated with plateletseither expressing PD-1 or loaded with cyclophosphamide.

2. Nucleic Acid and Amino Acid Sequences

In some embodiments, the cargo of the engineered platelets of theinvention may be a messenger RNA (mRNA). As used herein, the term“messenger RNA” (mRNA) refers to any polynucleotide which encodes apolypeptide of interest and which is capable of being translated toproduce the encoded polypeptide of interest in vitro, in vivo, in situor ex vivo. Such mRNA molecules may have the structural components orfeatures of any of those taught in International Publication No. WO2013/151666, which is incorporated herein by reference in its entirety.

c. CRISPR/Cas Systems

In some embodiments, a CRISPR/Cas gene editing system may be used toalter the genome of a megakaryocyte to produce the engineered plateletsdescribed herein. Alternatively, a CRISPR/Cas system may be packaged ina vesicle to be released on activation of the platelet by an antigenrecognized by the CPR CRISPR/Cas systems are bacterial adaptive immunesystems that utilize RNA-guided endonucleases to target specificsequences and degrade target nucleic acids. They have been adapted foruse in various applications in the field of genome editing and/ortranscription modulation. Any of the enzymes or orthologs known in theart or disclosed herein may be utilized in the methods herein for genomeediting.

In certain embodiments, the CRISPR/Cas system may be a Type IICRISPR/Cas9 system. Cas9 is an endonuclease that functions together witha trans-activating CRISPR RNA (tracrRNA) and a CRISPR RNA (crRNA) tocleave double stranded DNAs. The two RNAs can be engineered to form asingle-molecule guide RNA by connecting the 3′ end of the crRNA to the5′ end of tracrRNA with a linker loop. Jinek et al., Science,337(6096):816-821 (2012), which is hereby incorporated by reference inits entirety, showed that the CRISPR/Cas9 system is useful forRNA-programmable genome editing, and international patent application WO2013/176772 provides numerous examples and applications of theCRISPR/Cas endonuclease system for site-specific gene editing, which areincorporated herein by reference in their entirety. ExemplaryCRISPR/Cas9 systems include those derived from Streptococcus pyogenes,Streptococcus thermophilus, Neisseria meningitidis, Treponema denticola,Streptococcus aureas, and Francisella tularensis.

In certain embodiments, the CRISPR/Cas system may be a Type VCRISPR/Cpf1 system. Cpf1 is a single RNA-guided endonuclease that, incontrast to Type II systems, lacks tracrRNA. Cpf1 produces staggered DNAdouble-stranded break with a 4 or 5 nucleotide 5′ overhang. Zetsche etal. Cell. 2015 Oct. 22; 163(3):759-71, which is hereby incorporated byreference in its entirety, provides examples of Cpf1 endonuclease thatcan be used in genome editing applications, which is incorporated hereinby reference in its entirety. Exemplary CRISPR/Cpf1 systems includethose derived from Francisella tularensis, Acidaminococcus sp., andLachnospiraceae bacterium.

In certain embodiments, nickase variants of the CRISPR/Cas endonucleasesthat have one or the other nuclease domain inactivated may be used toincrease the specificity of CRISPR-mediated genome editing. Nickaseshave been shown to promote HDR versus NHEJ. HDR can be directed fromindividual Cas nickases or using pairs of nickases that flank the targetarea.

In certain embodiments, catalytically inactive CRISPR/Cas systems may beused to bind to target regions (e.g., gene encoding an antigen, such asa receptor) and interfere with their function. Cas nucleases such asCas9 and Cpf1 encompass two nuclease domains. Mutating critical residuesat the catalytic sites creates variants that only bind to target sitesbut do not result in cleavage.

In certain embodiments, a CRISPR/Cas system may include additionalfunctional domain(s) fused to the CRISPR/Cas endonuclease or enzyme. Thefunctional domains may be involved in processes including but notlimited to transcription activation, transcription repression, DNAmethylation, histone modification, and/or chromatin remodeling. Suchfunctional domains include but are not limited to a transcriptionalactivation domain (e.g., VP64 or KRAB, SID or SID4X), a transcriptionalrepressor, a recombinase, a transposase, a histone remodeler, a DNAmethyltransferase, a cryptochrome, a light inducible/controllable domainor a chemically inducible/controllable domain.

In certain embodiments, a CRISPR/Cas endonuclease or enzyme may beadministered to a cell or a patient as one or a combination of thefollowing: one or more polypeptides, one or more mRNAs encoding thepolypeptide, or one or more DNAs encoding the polypeptide.

d. Guide Nucleic Acids

In certain embodiments, guide nucleic acids may be used to direct theactivities of an associated CRISPR/Cas enzymes to a specific targetsequence within a target nucleic acid. Guide nucleic acids providetarget specificity to the guide nucleic acid and CRISPR/Cas complexes byvirtue of their association with the CRISPR/Cas enzymes, and the guidenucleic acids thus can direct the activity of the CRISPR/Cas enzymes.

In one aspect, guide nucleic acids may be RNA molecules. In one aspect,guide RNAs may be single-molecule guide RNAs. In one aspect, guide RNAsmay be chemically modified. In certain embodiments, more than one guideRNAs may be provided to mediate multiple CRISPR/Cas-mediated activitiesat different sites within the genome.

3. Small Molecules Drugs

In some embodiments, the cargo in the vesicles of an engineeredplatelets described herein is a small molecule drug such as, but notlimited to, (−)-Epigallocatechin 3-gallate, (−)-phenserine,(+)-calanolide A, (R)-folitixorin, (R)-mequitazine, (S)-pantoprazolesodium, [11C]DASB, [11C]-raclopride, [18F]FDG, [18F] HX4,1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea, 1,2-decanediol,11,11-di-deutero-ethyl linoleate, 11C-PBR-28, 123I-iometopane,124I-CLR-1404, 131I-MIBG, 131-Iodine, 13-cis-retinoic acid, 13C-labeledmethacetin, 13N-ammonia, 1400W94, 17 beta-estradiol, 17-alphahydroxyprogesterone caproate, 17-beta-estradiol, 17-beta-estradiolvalerate, 17-hydroxysteroid dehydrogenase inhibitors, 18F-EF5, 18F-FDG,2 L polyethylene glycol, 25-dihydroxy-vitamin D3, 25-OH vitamin D,2-chloroprocaine, 2-deoxyglucose, 2-Hydroxypropyl-Beta-Cyclodextrin,2MD, 2-methoxyestradiol, 4-aminopyridine, 4-aminosalicylic acid, 4-FEC,4-hydroxytamoxifen, 5-aminolevulinic acid, 5-aminosalicylic acid,5-aracytine, 5-fluorouracil (5-FU), 5-hydroxytryptophan,5-methoxypsoralen, 6-mercaptopurine, 6-thioguanine, 9-aminocamptothecin,9-aminofusin, 9-nitrocamptothecin, abacavir, abafungin, abametapir,abediterol, abexinostat, abiraterone, ABT-072, ABT-751, acadesine,acalabrutinib, acamprosate, acamprosate calcium, acarbose, acebilustat,acebutolol, aceclidine, aceclofenac, aceneuramic acid, acenocoumarol,Acetadote, acetaminophen, acetate-free bicarbonate, acetazolamide,acetic acid, acetylcholine, acetylcysteine, acetyl-L-carnitine,acetyl-L-carnitine hydrochloride, acetyl-L-leucine, acetylsalicylicacid, acetyl-salicylic acid, Acetylsalicylic acid (ASA), acetylsalicylicacid lysinate, aciclovir, acipimox, acitretin, aclarubicin, aclidinium,aclidinium bromide, acolbifene, acorafloxacin, acotiamide hydrochloride,ACP-104, acrivastine, ACT-01, ACT-280778, actinomycin D, acumapimod,acyline, adapalene, ADC-3680, Adderall XR, adefovir dipivoxil,ademetionine, adenosine, adinazolam, adipiplon, adomeglivant,adozelesin, adramycin, adrenalin, adrenaline, adriamycin, Advair, Advil,AE-941, afacifenacin fumarate, afatinib, afegostat, afeletecan,afimoxifene, aflibercept, aftobetin, afuresertib, aganepag isopropyl,agatolimod, agave inulin, agomelatine, Aiphagan, ajmaline, aladorian,alagebrium chloride, alanyl-glutamine dipeptide, albaconazole,albendazole, albiglutide, albitiazolium bromide, albumin, albuterol,albuterol sulphate, albuterpenoids, alcaftadine,alcipotriol/betamethasone, aldesleukin, aldoxorubicin, alectinib,aleglitazar, alemtuzumab, alendronate, alendronate sodium, alendronicacid, Alequel, Aleve, alphacalcidol, alfentanil, alfuzosin,algeldrate/magnesium oxide, Alimta, alisertib, aliskiren, alisporivir,alitretinoin, alizapride, allantoin, allisartan isoproxil,allopregnanolone, allopurinol, all-trans retinoic acid, almorexant,almotriptan, Alodan, alogliptin benzoate, alosetron, alovudine,alpelisib, alpha lipoic acid, alpha tocopherol, alpha-1 antitrypsin,alpha-cyclodextrin, alpha-glucosidase inhibitor, alpha-interferon,alpha-lipoic acid, alpha-tocopherol, alpha-tocopherol acetate,alpha-trichosanthin, alprazolam, alprostadil, alprostadil alphadex,ALS-08, altinicline, Altropane, aluminium MgS, aluminum hydroxide,alvespimycin hydrochloride, alvimopan, alvocidib, amantadine, amantadinehydrochloride, ambrisentan, ambroxol, ambroxol hydrochloride, AMD-070,amdoxovir, amelubant, amenamevir, Ametop, amfetamine, amibegron,amifampridine phosphate, amifostine, amikacin, amiloride, amiloridehydrochloride, amino acid, Aminocaproic Acid, aminoglutethimide,aminoguanidine, aminolevulinic acid, aminolevulinic acid hydrochloride,aminophylline, aminopterin, amiodarone, amiprilose, amiselimod,amisulpride, amitifadine hydrochloride, amitriptyline, Amitriptylinehydrochloride, amlexanox, amlodipine, amlodipine besilate, amlodipinebesylate, amlodipine camsylate, amlodipine maleate, ammonium lactate,amnion, amodiaquine, amonafide dihydrochloride, amonafide L-malate,amorolfine, amoxapine, amoxicillin, amoxicillin clavulanate, amoxicillinMR, amoxicillin/clavulanate, amoxicillin-clavulanic acid, amoxycillin,amphetamine, amphetamine aspartate, amphetamine sulphate, amphotericin,amphotericin B, ampicillin, ampicillin sodium,ampicillin/flucloxacillin, amprenavir, amrubicin, amsacrine,amsilarotene, AN-2898, AN-9, anacetrapib, anagliptin, anagrelide,anamorelin, anastrozole, anatibant, ancriviroc, ancrod, androgen,Androxy, anecortave, angiotensin converting enzyme inhibitor,angiotensin I, angiotensin II, Angiozyme, anidulafungin, aniracetam,annamycin, antazoline, anthocyanin, anthracycline, anti-emetic,antihistamine, antilymphocyte globulin, antineoplaston A-10,antineoplaston A10-I, antineoplaston AS2-1, Antioxidant Vitamins,antipsychotic, antiretroviral drugs, antithymocyte globulin,anti-thymocyte globulin, apabet, apadenoson, apaziquone, apelin,apheresis, apilimod, apimostinel, apitolisib, apixaban, aplaviroc,aplindore, apomorphine, Apovir, apratastat, apremilast, aprepitant,apricitabine, apricoxib, aprotinin, AR-623, Ara-C, arachidonic acid,aracytine, Aralast, aramchol, arasertaconazole, arbaclofen, arbaclofenplacarbil, arbekacin sulphate, arbutin, ARC-100, arformoterol,argatoroban, argatroban, arginine, arginine vasopressin, ARH-1,arhalofenate, arimoclomol, aripiprazole, armodafinil, arogliptin,arsenic trioxide, artefenomel mesylate, artemether,artemether-lumefantrine combination, artemisinin, artemisone, artemotil,artenimol, arterolane, arterolane maleate, artesunate,artesunate+mefloquine, artesunate-amodiaquine, articaine, articainehydrochloride, arundic acid, arzoxifene, asapiprant, ASCJ-9, ascorbate,ascorbic acid, asenapine, asimadoline, ASM-024, asoprisnil, aspirin,astaxanthin, astodrimer, asunaprevir, AT-101, ataciguat, atagabalin,ataluren, atamestane, atazanavir, atazanavir sulphate,atazanavir/ritonavir, atecegatran fexenetil, Atelvia, Atenativ,atenolol, atevirdine, ATHX-105, atiprimod, atiratecan, Ativan,atomoxetine, atopaxar, atorvastatin, atovaquone, atracurium, atracuriumbesylate, atrasentan, atreleuton, Atripla, atropine, auranofin,auriclosene, AVAC, avacopan, avagacestat, avanafil, avasimibe,avatrombopag, AVE-0657, AVE-2268, avibactam sodium, Avil, avobenzone,avoralstat, avosentan, AWD-12-281, axelopran, Axiron, axitinib,axomadol, azacitidine, azathioprine, AZD-1775, AZD-4547, AZD-9668,Azedra, azelaic acid, azelastine, azelastine hydrochloride, azeliragon,azelnidipine, azidothimidine, azilsartan, azilsartan medoxomilpotassium, azimilide, azithromycin, azithromycin dihydrate, azosemide,aztreonam, aztreonam lysine, bacitracin, baclofen, bafetinib, baicalin,balaglitazone, balicatib, balsalazide, bambuterol, banoxantrone,barasertib, bardoxolone methyl, baricitinib, barnidipine, basiliximab,basimglurant, basmisanil, batabulin, batefenterol succinate, bavisant,bazedoxifene, BCG vaccine, BCNU, becatecarin, beclabuvir, beclometasone,beclometasone dipropionate, beclomethasone, beclomethasone dipropionate,becocalcidiol, Beconase, bedaquiline, bedoradrine, bedrocon, belinostat,belladonna, belnacasan, beloranib, belotecan, bempedoic acid,benazepril, bendamustine, bendroflumethiazide, beneh, benfotiamine,benidipine, benserazide, bentamapimod, benzalkonium, benzalkoniumchloride, benzathine penicillin, benzbromarone, benznidazole,benzocaine, benzodiazepine, benzophenone-3, benzoyl peroxide,benztropine, benzydamine hydrochloride, benzylic alcohol,benzylpenicillin, benzylpiperazine, Bepantol, bepotastine, beractant,beraprost sodium, berberine, berubicin, besifloxacin, besifovir, betaerythropoietin, beta-1,3/1,6-glucan, beta-blocker, beta-blockers,beta-carotene, beta-cryptoxanthin, betadine, Betafectin, betahistine,betaine, Betaine anhydrous, beta-lactamase inhibitor, Betamarc,betamethasone, betamethasone dipropionate, betamethasone mousse,betamethasone valerate, betamethasone dipropionate, beta-tricalciumphosphate bone substitute, betaxolol, betaxolol hydrochloride,bethanechol, bethanechol chloride, betrixaban, betulinic acid,bevacizumab, bevenopran, bevirimat, bexagliflozin, bexarotene,bezafibrate, BF-derm1, BGP-15, BI-54903, biapenem, bicalutamide,bicifadine, bifeprunox, bifidobacterium, Bifidobacterium bifidum,Bifidobacterium infantis 35624, bifonazole, biguanide, BIB-021,bilastine, BILR-355-BS, bimatoprost, bimoclomol, bimosiamose, bindarit,binimetinib, binodenoson. Bio-25, biotin, biperiden, biphentin,birabresib dihydrate, biricodar, birinapant, bisacodyl, biskalcitratepotassium, bismuth, bismuth citrate, bismuth potassium citrate, bismuthsodium tartrate, bismuth subcitrate, bismuth subsalicylate, bisoprolol,bisoprolol fumarate, bisphosphate, bitopertin, bixalomer, bleomycin,bleomycin sulphate, blonanserin, BMP-7, BNC-105P, boceprevir, boricacid, boron-anticancers, bortezomib, bosentan, bosutinib, bradanicline,bradykinin, Bramitob, branched chain amino acid, brecanavir,brexpiprazole, Bricanyl, Bricasol, brimonidine, brimonidine tartrate,Brinavess, brinzolamide, brivanib alaninate, brivaracetam, brivudine,brolucizumab, bromfenac, bromfenac sodium, bromhexine, bromocriptine,brompheniramine, bronopol, brostallicin, brotizolam, bryostatin-1,BTI-320, BTL-TML-HSV, BTS-67582, bucindolol, budesonide,budesonide/formoterol, budesonine, budiodarone, bumetanide, bunazosin,buparlisib, bupivacaine, bupivacaine hydrochloride, bupivacaine withfentanyl, bupivacaine-clonidine, buprenorphine, buprenorphinehemiadipate hydrochloride, buprenorphine hydrochloride,buprenorphine/naloxone, bupropion, bupropion hydrochloride, bupropionSR, burapitant, burixafor, buserelin, buserelin acetate, buspirone,buspirone hydrochloride, busulfan, Busulfex, butalbital, butenafine,butoconazole, butoconazole nitrate, butorphanol, butorphanol tartrate,C5a, Cabaseril, cabazitaxel, cabergoline, cabotegravir, cabozantinibS-malate, Cacit D3, cadazolid, CAF regimen, caffeic acid, caffeine,caffeine citrate, caffeinol, Calcichew D3 Forte, calcipotriol,calcipotriol/betamethasone, calcitriol, calcium, calcium acetate,calcium ascorbate, calcium carbonate, Calcium chloride, calcium chloridedihydrate, calcium citrate, calcium dobesilate, calcium fluoride,calcium folinate, calcium glucarate, calcium gluconate, calciumhydrogenphosphate, calcium L-aspartate, calcium levofolinate, calciumphosphate, calcium polycarbophil, Calcium sodium phosphosilicate,Calcium supplements, calcium and vitamin D, calcium leucovorin,caldaret, calphactant, camicinal, camobucol, camptothecin,canagliflozin, candesartan, candesartan cilexetil, canertinib,canfosfamide, cangrelor, cannabidiol, Cannabidiol (CBD), cannabidivarin,cantharidin, capadenoson, capecitabine, capmatinib, Capolac,capravirine, Capros, capsaicin, captopril, carbamazepine, carbenoxolone,carbetimer, carbetocin, carbidopa, carbocisteine, carbocysteine,Carbogen, carbon [14C] oxaliplatin, carbon dioxide, carbon monoxide,carbondioxide, carboplatin, Carboxymethylcellulose sodium, cardidopa,cardonutrient, carfilzomib, carglumic acid, cariporide, cariprazine,carisbamate, carisoprodol, carmegliptin, carmoterol, carmustine,camitine, carotegrast methyl, carteolol, carteolol hydrochloride,carvedilol, carvedilol phosphate, CASAD, casein, casopitant,caspofungin, catechin, CBT-1, CCPI, cebranopadol, cediranib, cefaclor,cefadroxil, cefalexin, cefazolin, cefazolin sodium, cefdinir, cefditorenpivoxil, cefepime, cefilavancin, cefixime, cefmetazole, cefoperazone,cefotaxime, cefotetan, cefoxitin, cefozopran, cefpirome, cefpodoxime,cefprozil, ceftaroline, ceftaroline fosamil, ceftazidime, ceftibuten,ceftobiprole medocaril, ceftolozane sulphate, ceftriaxone, cefuroxime,celecoxib, Celestone, celgosivir, celiprolol, celivarone, Cenestin,cenicriviroc, cenobamate, censavudine, centanafadine, Centrum forte,Centrum Silver, cephalexin, cephalosporin, ceralifimod, ceramide,ceritinib, cerium nitrate, cerivastatin, cerlapirdine, certoparin,cetaphil, cethromycin, cetilistat, cetirizine, cetylpyridinium chloride,cevimeline, chenodeoxycholic acid, CHF-1535, CHF-5992, chitosan,chitosan-thiomer, chlorambucil, chloramphenicol, chlordiazepoxide,chlorhexidin, chlorhexidine, chlorhexidine digluconate, chlorhexidinegluconate, chlorhexidine-alcohol, chlorhydrate, chlormadinone acetate,chloroprocaine, chloroquine, chloroquine diphosphate, chloroquinephosphate, chlorpheniramine, chlorpheniramine maleate, chlorproguanil,chlorpromazine, chlortalidone, chlorthalidone, chlorzoxazone,cholecalciferol, cholecystokinin-8, cholesterol absorption inhibitors,cholestyramine, cholic acid, choline, choline alfoscerate, cholinediepalrestat, choline fenofibrate, choline magnesium trisalicylate,chondroitin, chondroitin sulphate, CHP-HER2, Chromium cobalt, ChromiumPicolinate, CHS-131, CHVP-interferon, ciclesonide, cicletanine,ciclopirox, ciclopirox olamine, ciclosporin, ciclosporine A, cidofovir,cilansetron, cilastatin, cilazapril, cilnidipine, cilomilast,cilostazol, ciluprevir, cimetidine, cimicoxib, cinacalcet, cinaciguat,cindunistat hydrochloride maleate, cinhyaluronate sodium, cinitapride,cinitapride tartrate, cinnamaldehyde, cinnarizine, cipargamin,ciprofibrate, ciprofloxacin, ciprofloxacin hydrochloride, ciraparantag,Cisapride, cisatracurium, cisatracurium besilate, cisplatin, cisplatinliposomal, cisplatinum, citalopram, citalopram hydrobromide, Citanest,citicoline, citrate, citrate fentanyl, citric acid, citric acidmonohydrate, citrulline, CK-2017357, cladribine, Clarinex,clarithromycin, clavulanate, clavulanate potassium, clavulanic acid,clazosentan, clebopride, clemastine, clemastine fumarate, clenbuterol,clevidipine, clevudine, clindamicin, clindamycin, clindamycin phosphate,clindamycin/benzoyl peroxide, Clinisol, clioquinol, clobazam,clobetasol, clobetasol propionate, Clobex, clodronic acid, clofarabine,clofazimine, clomethiazole edisylate, clomifen, clomifene, clomifenecitrate, clomiphene, clomiphene citrate, clomipramine, clonazepam,clonidine, clonidine hydrochloride, clopidogrel, clopidogrel hydrogensulphate, clopidogrel napadisilate, Cloratadd-D, clorazepate,Clostridium butyricum MIYAIRI 588, clotrimazole, cloxacillin, clozapine,coamoxiclav, cobalamin, cobamamide, cobicistat, cobimetinib,cobiprostone, cocaine, codeine, codeine phosphate, coenzyme Q10,Coeruleus, cogentin, Cogentus, colchicine, colcine, colecalciferol,colecalciferol D3, coleneuramide, colesevelam, colestilan, colestipol,colfosceril palmitate, colistimethate, colistimethate sodium, colistin,colistin sulphate, colistineb, colloidal bismuth, colloidal bismuthtartrate, coloxyl, coluracetam, combretastatin, Comp-01, Comp-02,Comp-03, Comp-04, conivaptan, conivaptan hydrochloride, conjugatedestrogen, conjugated estrogens, controlled-release carvedilol,copanlisib, copper, copper histidine, coQ10, cortexolone17alpha-propionate, corticosteroid, cortisol, cortivazol, cositecan,CosmoFer, cotrifazid, cotrimoxazol, cotrimoxazole, co-trimoxazole,COX-inhibitor, CPI-613, CRAd 3/5-delta, creatine, creatine ethyl ester,creatine monohydrate, crenolanib, crisaborole, crizotinib, CRM-197,crobenetine, crofelemer, cromoglicate, cromoglicic acid, cromolynsodium, Crsytalloids, C-Tb, CTO, CUDC-305, Curcuma aeruginosa, curcumin,curcuminoids, curdlan sulphate, cutamesine dihydrochloride, CX-516,cyanocobalamin, cyclizine, cyclizine lactate, cyclobenzaprine,cyclobenzaprine hydrochloride, cyclodextrin, cyclodextrin-combineddiclofenac. Cyclogest, cyclopentolate, cyclophophamide,cyclophosphamide, cyclophosphamide monohydrate, cyclophosphan,cycloserine, cyclosporin, cyclosporine, cyclosporine A, cyclosporinemicroemulsion, cyclphosphamide, cyproheptadine, cyproterane acetate,cyproteron, cyproterone, cyproterone acetate, cysteamine, Cysteaminehydrochloride, cysteine, cysteine hydrochloride monohydrate, cytarabine,cytarabine arabinoside, cytarabine-asparagine conjugate, cytophosphan,Cytosin-Arabinosid, cytosine arabinoside, cytoxan, Cytozar, D3 vitamine,DA-9601, dabigatran etexilate, dabrafenib, dacarbazine, daclatasvir,daclizumab, dacomitinib, dactinomycin, dactolisib, daglutril, daidzein,dalbavancin, dalcetrapib, dalfopristin, dalteparin sodium,D-amphetamine, danazol, danirixin, danoprevir, dantrolene, dantrolenesodium, danusertib, dapaconazole, dapagliflozin, dapagliflozinpropanediol, dapansutrile, dapivirine, daporinad, dapoxetine, dapsone,dapsone gel, darapladib, darifenacin, darinaparsin, darolutamide,darotropium bromide, darunavir, darunavir/ritonavir, darusentan,dasabuvir, dasatinib, dasotraline, daunorubicin, daunorubicinhydrochloride, daunorubicine, D-cycloserine, dDAVP, DDP, DE-104, DE-110,DE-112, Deanxit, Debio-1450, Debio-1452, decadron, decarbazine,decernotinib, decitabine, decoglurant, Decuprate, defactinib,deferasirox, deferiprone, deferitazole, deferoxamine, deferoxaminemesylate, deflazacort, dehydroepiandrosterone, delafloxacin, delamanid,delanzomib, delapril, delapril hydrochloride, delavirdine, Delazine,deleobuvir, deligoparin sodium, delorazepam, delta-8-THC,delta-9-tetrahydrocannabinol, denagliptin, denufosol tetrasodium,Depacon, Depade, depo-medroxyprogesterone, depomedroxyprogesteroneacetate, depomethylprednisolone, depotestosterone, DER-45-EV,derenofylline, dersalazine sodium, desferrioxamine, desflurane,desipramine, desloratadine, desmopressin, desmopressin acetate,desogestrel, desonide, desote fumarate, desoximetasone, desvenlafaxine,Detox-B adjuvant, deutetrabenazine, dexamethasone, dexamethasoneacetate, dexamethasone cipecilate, dexamethasone diphosphate,dexamethasone phosphate, dexamethasone sodium phosphate, dexamfetamine,dexanabinol, dexchlorpheniramine, dexedrine, dexelvucitabine,dexfenfluramine, dexibuprofen. Dexid, dexisometheptene mucate,dexketoprofen, dexketoprofen trometamol, dexlansoprazole, dexlipotam,dexioxiglumide, dexmecamylamine, dexmedetomidine, dexmethylphenidate,dexniguldipine, dexpanthenol, dexpramipexole, dexrazoxane, dexrazoxanehydrochloride, dextofisopam, dextran, dextroamphetamine,dextroamphetamine saccharate, dextroamphetamine sulphate,dextromethorphan, dextromethorphan hydrobromide, dextropropoxyphene,dextropropoxyphene hydrochloride, dextrose, dexverapamil, dezocine,DHEA, diacerein, diacetylmorphine, Dialysate calcium, Diamel,diammindichloridoplatin, diamorphine, diamorphine hydrochloride,dianhydrogalactitol, dianicline, Diao Xin Xue Kang, diazemuls, diazepam,diazepam autoinjector, diazoxide, diazoxide choline, dibasic dihydratesodium phosphate, dibasic sodium phosphate, dibekacin, dichlorphenamide,Diclazuril, diclofenac, diclofenac diethylamine, diclofenac potassium,diclofenac sodium, dicloxacillin, didanosine, dienogest,diethylcarbamazine, diethylnorspermine, diethylpropion,diethylstilbestrol, diflomotecan, diflunisal, difluprednate, digitoxin,digoxin, dihematoporphyrin, dihomo gamma-linolenic acid, dihydralazine,dihydroartemisinin, dihydroartemisinin-piperaquine, dihydrocodeine,dihydroergotamine, dihydroergotamine mesylate, dihydroxy vitamin D3,diiodothyropropionic acid and its analogs, diiphenhydramine, dilaudid,dilmapimod, diltiazem, diltiazem hydrochloride, dimenhidrinate,dimenhydrinate, dimesna, dimethindene, dimethindene maleate, dimethylfumarate, dimethylfumarate, dimiracetam, dinoprostone, diosmin,diphencyprone, diphenhydramine, diphenylcyclopropenone, dipirone,dipraglurant-IR, dipyradimole, dipyridamole, dipyrone, diquafosolsodium, diquafosol tetrasodium, disopyramide, Dispase H, disufentonsodium, disulfiram, dithranol, ditiocarb sodium, DLBS-1033, DLBS-1425,D-methadone, DNE3, dobutamine, docetaxel, dociparstat, doconexent,doconexent ethyl ester, docosahexaenoic acid [DHA], docosahexaenoic acidmonoglycerides, docosanol, docusate, docusate sodium, dofetilide,dolasetron, dolastatin-10, dologesic, dolutegravir, domperidone,donepezil, donepezil hydrochloride, donu, dopamine, dopexamine,doramapimod, doravirine, doripenem, dorzolamide, dorzolamidehydrochloride, dorzolamide hydrochloride+timolol maleate, dothiepin,dovitinib, doxapram, doxazosin, doxazosin mesylate, doxepin, doxepinhydrochloride, doxercalciferol, doxifluridine, Doxil, doxophylline,doxorubicin, doxorubicin etarfolatide, doxorubicin HCl liposome,doxorubicin hydrochloride, doxorubicin hydrochloride liposome,doxycycline, doxycycline hyclate, doxylamine, doxylamine succinate,D-penicillamine, DPP-IV inhibitors, DPS-102, draflazine, drinabant,dronabinol, dronedarone, droperidol, dropropizine, drospirenone,drotaverine, droxidopa, D-tagatose, D-TRANS fentanyl, Duac, dual-releasehydrocortisone, dulaglutide, Dulcolax, duloxetine, Duracain, duramorph,Durolane, dutasteride, dutogliptin, duvelisib, duvoglustat, D-xylose,dydrogesterone, dyhydroprogesterone, DZ-1, E. coli Nissle, E-7016,E-7820, ebastine, ebselen, EC-17, ecabet, ecabet sodium, Echinacea,econazole nitrate, ecopipam, ecosprin, ecraprost, Ecural, edaglitazone,edaravone, edetate calcium disodium, edivoxetine, edonerpic maleate,edotecarin, edoxaban, EES0000645/A, efaproxiral, efatutazone, efavirenz,efinaconazole, eflornithine, efonidipine hydrochloride, EGb-761, EGCG,eicosapentaenoic acid, eicosapentanoeic acid, elacestrant, elacridar,elacytarabine, elafibranor, elagolix, elamipretide, elbasvir, elbion,eldecalcitol, eleclazine, elesclomol sodium, eletriptan, eliglustattartrate, elinogrel, eliprodil, ELND-005, elobixibat, elocalcitol,Elomet, Elosalic, elsamitrucin, eltoprazine, eltrombopag, elubrixin,eluxadoline dihydrochloride, elvitegravir, elvorin, elvucitabine,elzasonan, Emdogain, emedastine, emepepimut-S, emicerfont, emivirine,emixustat, empagliflozin, emricasan, emtricitabine, enalapril, enalaprilmaleate, enalaprilat, enasidenib, encaleret sulphate, enclomifenecitrate, enclomiphene, encorafenib, endocannabinoidpalmitoylethanolamide, endonase, endotoxin, endoxan, enecadin,enflurane, enfuvirtide, eniluracil, ENMD-2076, enobosarm, enocitabine,enoxaparin sodium, enoximone, entacapone, entecavir, entecavir maleate,enteric-coated mycophenolate sodium, enteric-coated tegafur-uracil,Enteroaggregative E. coli, entinostat, entonox, enzalutamide,enzastaurin, epacadostat, Epadel, epairestat, eperisone, eperisonehydrochloride, epetirimod, epetraborole, ephedrine, Epiceram,epidoxirubicin, epidoxorubicin, epidural/paravertebral analgesia,epigallocatechin gallate, epigallocatechin-3-gallate,epigallocatechin-gallate, Epiggallocatechin, epinastine, epinastinehydrochloride, epinephrine, epirubicin, epirubicin hydrochloride,Episalvan, eplerenone, eplivanserin, epoprostenol, Eppikajutsutou,eprodisate, eprosartan, eprotirome, epsilon-aminocaproic acid,eptastigmine, eptifibatide, Equine antithymocyte immunoglobulin,Equisetum arvense, eravacycline, erdosteine, Eremostachys laciniata,ergocalciferol, ergotamine, eribaxaban, eribulin mesylate, Eritex,eritoran, erlotinib, ertapenem, erteberel, ertugliflozin, erythromycin,erythromycin lactobionate, erythropoetin, erythropoietin beta,ESAT-6CFP10, esaxerenone, Escherichia coli endotoxin, escitalopram,esflurbiprofen, Eskalith, esketamine, esketamine hydrochloride,eslicarbazepine acetate, Esmeron, esmirtazapine, esmolol, esmololhydrochloride, esomeprazole, esoxybutynin, esreboxetine, estazolam,estetrol, estradiol, estradiol acetate, estradiol cypionate, estradiolvalerate, estradiol/norethindrone acetate, estramustine, estramustinephosphate, estramustine phosphate sodium, Estratest, estriol, estriolE3, estrodiol, estrogen, estrogens, eszopiclone, etacrynic acid,etalocib, Etalpha, etanercept biosimilar, etazolate, Ethacrynic acid,ethambutol, ethambutol hydrochloride, ethanol, ethinyl estradiol,ethinyl estradiol/levonorgestrel, ethinylestradiol, ethiodized oil,ethosuximide, ethyl chloride, ethyl eicosapentaenoate, ethyl hydrogenfumarate calcium, ethyl hydrogen fumarate magnesium, ethyl hydrogenfumarate zinc, ethylenediaminetetraacetate, ethylhexyl triazone,ethynylcytidine, etidronic acid, etilefrine, etodolac, etomidate,etomidate Lipuro, etomoxir, etonogestrel, etonox, etoposide, etoposidephosphate, etoricoxib, etravirine, EV-06, evacetrapib, evatanepag,everolimus, eviprostat, evofosfamide, evogliptin, exatecan, exemestane,exenatide, exeporfinium chloride, exisulind, ezatiostat, ezetimibe,ezetimibe/atorvastatin, F-0434, F0-MO, F0-M1700, F160-M0, F160-M1000,F160-M1700, F80-M1000, F80-M1700, FA, facinicline hydrochloride,faldaprevir, famciclovir, famitinib L-malate, famotidine, fampridine,Fangji, farampator, farglitazar, faropenem, faropenem medoxomil,fasiglifam hemihydrate, fasitibant chloride, fasudil, favipiravir,FBG-18, FBP peptides, FE[50]C, FE[75]C, Fe-58, febuxostat, fedovapagon,fedratinib, felbamate, felbinac, felodipine, fenatnyl, fenobam,fenofibrate, fenoldopam, fenoterol, fenoterol prednisone, fenretinide,fentanyl, fentanyl citrate, fermagate, ferric carboxymaltose, ferriccitrate, ferric maltol, ferric pyrophosphate, Ferripel-3, ferroquine,ferrous fumarate, ferrous sulphate, ferumoxtran-10, ferumoxytol, FeSO4,fesoterodine fumarate, fevipiprant, fexinidazole, fexofenadine,fiboflapon, fibrinogen, fidaxomicin, filanesib, filgotinib, filgrastim,filibuvir, filorexant, fimaporfin, fimasartan, finafloxacin,finafloxacin hydrochloride, finasteride, finerenone, fingolimod,Fioricet, fipamezole, fish oil (eicosapentaenoic acid [EPA] plusdocosahexaenoic acid [DHA]), Fisiogel, fispemifene, Flagyl, Flavan-3-ol,flavanone, flavoxate, flecainide, flibanserin, flomoxef, flomoxefsodium, flopristin, florbenazine, florbetapir, flortaucipir F 18,flourinic acid, flovagatran, floxuridine, fluciclatide F 18,flucloxacillin, fluconazole, flucytosine, fludarabine, fludeoxyglucose,fludeoxyglucose F 18, fludrocortisone, flumazenil, flunarizine,flunisolide, fluocinolone acetonide, fluocinonide, fluorescein,fluorometholone, fluorometholone acetate, fluoropyrimidine,fluoroquinolones, fluorouracil, fluoxetine, fluoxetine hydrochloride,flupenthixol, flupentixol, fluphenazine, flupirtine, flurbiprofen,flurbiprofen axetil, flurbiprofen sodium, flutamide, fluticasone,fluticasone furoate, fluticasone propionate, flutrimazole, fluvastatin,fluvoxamine, FM-VP4, folacin, folate, folate/iron, FOLFIRI, FOLFOX4,FOLFOXIRI, folic acid, folinate, folinic acid, folitixorin calcium,follitropin beta, fonadelpar, fondaparinux sodium, Foradil, foretinib,formoterol, formoterol fumarate, forodesine, foropafant, fosalvudinetidoxil, fosamprenavir, fosamprenavir calcium, fosaprepitant,fosbretabulin, fosbretabulin disodium, foscarnet, foscarnet sodium,fosdagrocorat, fosdevirine, fosfluridine tidoxil, fosfomycin, fosfomycintrometamol, fosfructose, fosinopril, fosmidomycin, fosphenytoin,fospropofol, fostamatinib, fostemsavir tromethamine, Fostimone, Fostrap,fotemustine, fozivudine tidoxil, freselestat, Fresubin, frovatriptan,fructose, fructose-1,6-diphosphate, fruquintinib, frusemide, fucoidan,fulvestrant, fumarate, funapide, furaprevir, furazolidone, furosemide,fusidate sodium, fusidic acid, Fuzheng Huayu, gabapentin, gabapentinenacarbil, gaboxadol, gadobenic acid, gadobutrol, gadofosveset,gadolinium, gadopentetate dimeglumine, gadoterate meglumine,gadoversetamide, gadoxetate disodium, Galactooligosaccharide,Galacto-oligosaccharides, galantamine, galantamine CR, galeterone,gallium maltolate, gallium nitrate, gallopamil, gambogic acid,Gamma-Linolenic Acid, gamma-tocopherol, ganaxolone, ganciclovir,ganciclovir phosphonate, ganetespib, ganirelix acetate, ganstigmine,garenoxacin, garlic, gatifloxacin, GCS-100, G-CSF, gedatolisib,gefitinib, gelatin, Gelofusine, Gelpart, gemcabene, gemcitabine,gemcitabine elaidate, gemcitabine prodrug, gemfibrozil, gemifloxacin,gemigliptin, gemigliptin tartaric acid, Gemzar, Genaera, genistein,genistein+decitabine, gentamicin, gentamicin sulphate, gentamycin,gepirone, gepotidacin, gestodene, gestodone, gestrinone, gilteritinib,gimatecan, gimeracil, Ginkgo biloba, ginkgolides meglumine, ginsenosideRg3, ginsenoside-Rd, giripladib, gisadenafil, givinostat, GKT-831,glatiramer acetate, glecaprevir, glesatinib glycolate, glibenclamide,gliclazide, glimepiride, glinide, glipizide, glitazone, GLP-1 analog,glucagon-like peptide-1, glucocorticoids, glucocorticosteroid,glucosamine, glucosamine hydrochloride, glucose, glufosfamide, glutamicacid, glutamine, glutathione, glycerin, glycerol, glycerolphenylbutyrate, glycoprotein IIb/IIIa inhibitor, glycopyrolate,glycopyrrolate, glycopyrronium bromide, glycopyrronium tosylate,glycopyrroniumbromide, glycyrrhizin, glyminox, GM1, GM-CT-01, GnRHantagonist, gold sodium thiosulphate, golotimod, golvatinib tartrate,gonadotopin, gonadotropin, gonadotropins, GoodBelly probiotic, goserelinacetate, goshajinkigan, gosogliptin, gp100, GPO-Vir Z30, granisetron,granotapide, grazoprevir, grepafloxacin, griseofulvin, GR-MD-02,GSK-2269557, GSK-2330672, GSK-2339345, guaifenesin, guanethidine,guanfacine, Guizhi fulings, gusperimus trihydrochloride, GWP-42004,gycerol, Gynostemma pentapyllum, H2 blockers, haldol, halofuginone,haloperidol, haloperidol decanoate, halothane, Hangeshashin-to, HBW,HE-3286, Healon, Helico DR, heliox, heme arginate, hemin, hemoximer,heparin, heparin sodium, Her-2/neu, heroin, hexachlorophene,hexaminolevulinate hydrochloride, Hextend, HF-0220, Hibiscus sabdariffa,hidrotalcid, himantane, histamine dihydrochloride, homatropine, honey,hpFSH, HPP-404, HQK-1004, huachansu, Huai Qi Huang, human chorionicgonadotrophin, huperzine A, Hyabest J, hyaluronan, hyaluronate sodium,hyaluronic acid, hyaluronic acid hydrogel, hydralazine, hydrochloricacid, hydrochlorothiazide, hydrochlorothiazide tablet,hydrochlorthiazide, hydrocodone, hydrocodone bitartrate,hydrocodone/acetaminophen, hydrocortisone, hydrocortisone sodiumsuccinate, hydrocortisone-17-butyrate, hydrocortone, hydrogel, hydrogenperoxide, hydromorphine, hydromorphone, hydromorphone hydrochloride,hydroquinidine, hydroquinone, hydroxocobalamin, hydroxycarbamide,hydroxychloroquine, hydroxydaunorubicin, hydroxyethyl starch,hydroxyethylstarch solution, Hydroxyl-propyl-methyl cellulose powder,hydroxymethylbutyrate, hydroxynortriptyline, hydroxyprogesteronecaproate, hydroxypropyl cellulose, hydroxytryptophan, hydroxyurea,hydroxyzine, hylastan, Hylenex recombinant, hyoscine butylbromide,hyoscine hydrobromide, hyoscine N-butylbromide, hyoscyamine sulphate,hyperbaric bupivacaine, hypericin, Hypericum perforatum, hyperosmolardextrose, hypertonic saline, hypromellose, ibandronate, ibandronic acid,iberogast, iberogast N, IBH-B, ibipinabant, ibodutant, ibopamine,ibrutinib, ibudilast, ibuprofen, ibutamoren mesylate, ibuterol,ibutilide, icaritin, icodextrin, icofungipen, icosabutate, icosapent,icosapent ethyl, icosapent ethyl ester, icotinib hydrochloride,idalopirdine, idarubicin, idazoxan, IdB-1016, idebenone, idelalisib,idoxuridine, idrabiotaparinux sodium, idraparinux sodium, idronoxil,iferanserin, ifetroban, ifetroban sodium, IFN-alpha2b, ifosfamide,iguratimod, IHBG-10, IL-2, ilaprazole, ilepatril, iloperidone, iloprost,iloprost betadex clathrate, imagabalin, imatinib, ImCOOH, imeglimin,imexon, imidafenacin, imidapril, imiglitazar, imipenem, imipramine,imiquimod, imisopasem manganese, IMO-2125, implitapide, incyclinide,indacaterol, indacaterol acetate, indacaterol maleate, indacaterolxinafoate, indantadol, indapamide, indapamide SR, indeglitazar,indinavir, Indinol Forto, indiplon, indisetron, indisulam, Indium In 111anti-CD66 monoclonal antibody BW250/183, indocyanine green, indometacin,indomethacin, indoramin, industrial nitric oxide, inecalcitol,INF-alpha, infigratinib, infliximab, Ingavirin, ingenol mebutate,inhaled sodium nitrite, iniparib, injectable progestin, inosine, inosinepranobex, inositol, INS-1, insulin, insulin glargine, insulin NPH,intepirdine, interferon, interferon alpha, interferon alpha-2a,interferon alpha-2b, interferon beta-1a, interferon beta1b, interleukin,interleukin-2, interleukin-6, intetumumab, Intracel, intranasalketamine, intravenous immunoglobulin, intravenous minocycline,iobenguane I-131, iodine, iodine I 131 ethiodized oil, iodine I 131monoclonal antibody BC8, iodine tincture, iodixanol, iohexol, iopamidol,iopromide, ipragliflozin, ipratropium, ipratropium bromide, IPX-159,IPX-231, irbesartan, irinotecan, irinotecan hydrochloride, irinotecansucrosofate, irofulven, iron, iron folic acid, iron hydroxidepolymaltose, iron oxide, iron proteinsuccinylate, iron solution, ironsucrose, iron supplements, irosustat, irsogladine maleate, IRX-5183,ISA-51, isavuconazonium chloride/sulphate, Iscar, iseganan, Isobide,isocaloric diet, isocarboxazid, isoflavone, isoflavones, isoflurane,Isolyte-S, isoniazid, isoniazide, isoprinosine, isopropyl alcohol,isopropyl unoprostone, isoproterenol, isoquercetin, isosorbidedinitrate, isosorbide mononitrate, isosorbide-5-mononitrate, isosulfanblue, isotretinoin, isovaleramide, ispinesib, ispronicline, isradipine,israpafant, istaroxime, itacitinib, itasetron, itopride hydrochloride,itraconazole, itriglumide, ivabradine, ivabradine hydrochloride,ivacaftor, ivermectin, ixabepilone, ixazomib citrate, Jin Fu Kang,JNJ-56914845, Jobelyn, josamycin, Juglans regia extract, Juvidex,Kamikihi-to, kanamycin, kava, KD-018, Keflin, Kenalog, Kenalog-10,ketamine, ketamine hydrochloride, ketanserin, ketoconazole, ketoprofen,ketorolac, ketorolac tromethamine, ketotifen, KLH, Krestin, KRX-0402,KT6-971, KW-2450, KW-2478, KWA-0711, KX2-391, L9NC, labetalol,labradimil, lacidipine, Lacidofil Strong, lacosamide, lactated ringer'ssolution, lactic acid, LACTIN-V, lactitol, lactobacillus, Lactobacillusacidophilus, Lactobacillus acidophilus KS400, Lactobacillus caseirhamnosus, Lactobacillus delbrueckii, Lactobacillus paracasei F19,Lactobacillus paracasei LP-33, Lactobacillus plantarum 299v,Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus Sporogens,lactose, lactose monohydrate, lactulose, ladarixin, ladostigil,lafutidine, L-alanosine, lamivudine, lamotrigine, landiolol, lanicemine,laninamivir octanoate, laniquidar, lanoconazole, lanopepden,lanperisone, lansoprazole, lanthanum carbonate, lapaquistat, lapatinib,laquinimod, L-arginine, laromustine, laropiprant, larotaxel, L-ascorbicacid, lasmiditan, lasofoxifene, L-asparaginase, latanoprost,latanoprost, latanoprostene bunod, latrepirdine, lauric acid,lazabemide, LC-150444, L-carnitine, L-citrulline, LCL-161, Lcr-35,L-dopa, lecovorin, lecozotan, lecozotan SR, lederfolin, ledipasvir,lefamulin, leflunomide, lemborexant, lemuteporfin, lenalidomide,lenograstim, lentinan, lentinan viral, lenvatinib mesylate, LEO-80122,L-epinephrine, lercanidipine, lersivirine, lesinurad, lesogaberan,lestaurtinib, letaxaban, leteprinim, letermovir, letrozole, leucine,leucoverin, leucovorin, leucovorin calcium, leukapheresis, leukotrieneB4 (LTB4), leuprorelide, leuprorelin acetate, levalbuterol, levalbuterolhydrochloride, levamfetamine, levamisole, levamlodipine, levamlodipinebesylate, levetiracetam, levobetaxolol, levobupivacaine, levocabastinehydrochloride, levocarnitine, levocetirizine, levocetirizinedihydrochloride, levodopa, levofloxacin, levofolinate, levogestrel,levoketoconazole, levoleucovorin, levo-leucovorin, levomequitazine,levomilnacipran, levonorgestrel, levo-phencynonate hydrochloride,levorphanol, levosalbutamol, levosimendan, levosulpiride, levothyroxine,levothyroxine sodium, levotofisopam, lexibulin, lexipafant, L-folinicacid, L-glutamine, LH-RH agonist, liafensine, liarozole, Libifem,licarbazepine, licochalcone A, licofelone, lidocaine, lidocainechlorhydrate, lidocaine-prilocaine, lifibrol, lifitegrast, lignocaine,LIK-066, limaprost, Limtop, linagliptin, linaprazan, lincomycin,linezolid, linifanib, linoleic acid, linopristin, linsitinib,liothyronine, liothyronine sodium, lipid, lipiodol, Lipiodol-ethanolmixture, Lipocine, LipoCol, lipoic acid, lipopolysaccharide, liposomalamphotericin B, liposomal cisplatin, liposomal doxorubicin, liposomalpaclitaxel, liposomal prostaglandin E-1, liposomal vincristine, LiprocaDepot, lisavanbulin hydrochloride, lisdexamfetamine, lisinopril,lisofylline, lisuride, lithium, lithium carbonate, Lithium citrate,lithium salt, litronesib, lixivaptan, L-leucovorin, L-leucyl-L-leucinemethyl ester, L-NMMA, lobaplatin, lobeglitazone, lobeline, lobelinesulphate, lobucavir, lodenafil carbonate, lodenosine, lofepramine,lofexidine, lomibuvir, lomitapide, lomustine, Lomustine (CCNU),lonafarnib, lonaprisan, Long chain fatty acids, long-chainpolyunsaturated fatty acids, lonidamine, loperamide, loperamidehydrochloride, loperamide oxide, lopinavir, lopinavir/ritonavir,loratadine, lorazepam, lorcaserin, lorediplon, lormetazepam, L-ornithineL-aspartate, lornoxicam, losartan, losartan potassium, losigamone,losmapimod, loteprednol etabonate, lovastatin, loxapine, loxoprofen,lubiprostone, lucanthone, lucitanib hydrochloride, luliconazole,lumacaftor, lumateperone toluenesulfonate, lumefantrine, lumicitabine,luminespib, lumiracoxib, lunacalcipol, lurasidone, lurbinectedin,Lurotin, lurtotecan, luseogliflozin hydrate, lusutrombopag, lutein,LY-2090314, LY-2623091, Lybrido, lycopene, lymecycline, lynestrenol,Lysomucil, macimorelin, macitentan, Macrolids, mafenide, mafosfamide,MAGE-A1, MAG-EPA oil, magnesium, magnesium aluminum hydroxide, magnesiumcarbonate, magnesium chloride, magnesium chloride hexahydrate, magnesiumcitrate, magnesium gluconate, magnesium hydroxide, magnesiumisoglycyrrhizinate, magnesium oxide, magnesium sulphate, magnesiumvalproate, malathion, managlinat dialanetil, mangafodipir, manganese,manidipine, manidipine dihydrochloride, manitimus, mannitol, mapracorat,maprotiline, maralixibat chloride, maraviroc, maribavir, Marijuana,marimastat, marvelone, masilukast, masitinib, masoprocol, mavoglurant,maxacalcitol, mazindol, MCC-135, MDR1, MDT-10013, mebendazole,mebeverine, mebeverine hydrochloride, mecamylamine, mechlorethamine,meclinertant, meclizine, mecobalamin, mecobalamin monohydrate, Mediumchain fatty acids, medroxyprogesterone, medroxyprogesterone acetate,mefenamic acid, mefloquine, megestrol, megestrol acetate, meglumineantimoniate, melagatran, Melan-A, melarsoprol, melatonin, meldonium,melfalan, meloxicam, melperone, melphalan, melphalan hydrochloride,memantine hydrochloride, menaquinone, menaquinone-7, menatetrenone,Meniace, menotropin, menotropins, menstrogol, mepacrine, meperidine,mephalan, mepivacaine, mepivacaine chlorhydrate, mepivacainehydrochloride, mepridine, MER-104, merbarone, mercaptamine, mercaptaminebitartrate, mercaptopurine, mericitabine, merimepodib, meropenem,mesalamine, mesalazine, mesna, metadoxine, Metafolin, Metaglip,metamizole, metamizole sodium, metaraminol, Meteospasmyl, metformin,metformin glycinate, metformin HCl, metformin hydrochloride, metforminSR, methacholine, methadone, methadone hydrochloride, methamphetamine,methazolamide, methimazole, methionine, methocarbamol, methohexital,methotrexate, methotrimeprazine, methoxsalen, methoxyflurane,methoxypsoralen, methyl aminolevulinate hydrochloride, methylprednisolone, methyl prednisolone acetate, methylcobalamin,methyldibromoglutaronitrile, methyldopa, methylene blue,methylnaltrexone bromide, methylphenidate, methylphenidatehydrochloride, methylprednisolone, methylprednisolone aceponate,methylprednisolone acetate, methylprednisolone sodium succinate,methylprednisone, methylsamidorphan, methylsulfonylmethane,methyltetrahydrofolate, methylthioninium chloride, metirosine,Metobes-compound, metocloperamide, metoclopramide, metolazone,metoprolol, metoprolol succinate, metoprolol tartrate, metoprolol XL,metranidazole, MetroGel, metronidazole, metronomic cyclophosphamide,metyrapone, mexiletine, mexiletine hydrochloride, Mexoryl SX, MexorylX1+titanium dioxide, mianserin, mibampator, MIBG, miconazole, miconazolenitrate, microalgal oil, micronized progesterone, micronutrient mixture,midazolam, midazolam hydrochloride, middle-chain and polyunsaturatedfatty acids, midodrine, midostaurin, mifepristone, miflonide,migalastat, miglitol, miglustat, milataxel, milnacipran, milrinone,miltefosine, milveterol, mimopezil, minocycline, minocyclinehydrochloride, minodronic acid, minoxidil, mirabegron, miriplatinhydrate, mirodenafil, mirtazapine, misoprostol, mitemcinal, mitiglinide,mitoguazone, mitolactol, mitomycin, Mitomycin C, mitoquinone/mitoquinolredox mixture, mitotane, mitoxantrone, mivacurium, mivacurium chloride,mivobulin, mixed salt amphetamine, mizolastine, mizoribine, MK-0782,MK-0893, MK-2206, MK-7622, MK-8457, MMF, mocetinostat dihydrobromide,moclobemide, modafinil, moexipril, molidustat, molindone, molsidomine,molybdenum, momelotinib, mometasone, mometasone furoate, monolaurin,monosodium glutamate, montanide ISA-51, montelukast, montelukast sodium,moracizine, morphine, morphine chloride, morphine glucuronide, morphinehydrochloride, morphine sulphate, mosapride, motesanib diphosphate,motexafin gadolinium, motexafin lutetium, motolimod, moxidectin,moxifloxacin, moxifloxacin hydrochloride, moxonidine, MP-435,MSC-apceth-111, MT-102, Mucaine, Mucopolysaccharide, Mucuna pruriens,multivitamins, muparfostat sodium, mupirocin, muraglitazar, mustine,Mycobacterium w, mycophenolate acid, mycophenolate mofetil,mycophenolate sodium, mycophenolic acid, mycostatin, Mydriasert,Myfenax, Myrtus Communis L, mytomycin, N2O-O2, nabilone, nabiximols,nabumetone, N-acetyl cysteine, N-acetylcisteine, N-acetylcysteine,N-acetyl-L-cysteine, N-acetyl-p-aminophenol, NaCl, nadifloxacin,nadolol, nadroparin calcium, nafamostat, nafamostat mesilate, nafarelin,NaFeEDTA, naftifine, naftifine hydrochloride, naftopidil, nalbuphine,nalbuphine sebacate, naldemedine, nalfurafine, nalmefene, naloxegol,naloxone, naltrexone, naltrexone hydrochloride,naltrexone-poly(DL-lactide), naluzotan, Namodenoson, nandrolone,Naoxintong, napabucasin, naphthoquine, naproxcinod, naproxen, naproxenetemesil, naproxen sodium, naratriptan, naronapride, narrowband UVB,nasapaque, nastorazepide calcium, natamycin, nateglinide, navamepent,navarixin, naveglitazar, navitoclax, N-chlorotaurine, nebentan,nebicapone, nebivolol, nebulized amikacin, nebulized budesonide,Nebusal, nedaplatin, nefazodone, nefiracetam, neflamapimod, nefopam,neladenoson bialanate, nelarabine, nelfinavir, nelivaptan, nelociguat,nelonicline, nelotanserin, nemonoxacin, nemorubicin, neomycin, neomycinsulphate, neostigmine, nepadutant, nepafenac, nepicastat, neramexane,neratinib, neridronic acid, nerispirdine, netarsudil, netilmicin,netivudine, netupitant, Neu-120, neurotropin, nevirapine, niacin,niacinamide, Niacor, NIC5-15, nicardipine, nicergoline, Niclosamide,niconinamide, nicorandil, nicotinamide, nicotine, nicotine polacrilex,nicotinic acid, nifedipine, nifurtimox, Nigella sativa, nikkomycin Z,nilotinib, nilutamide, nilvadipine, nimesulide, nimodipine, nimorazole,nimotuzumab, nimustine hydrochloride, nintedanib, nipradilol, niraparib,nirogacestat, nitazoxanide, nitisinone, nitrazepam, nitrendipine, nitricoxide, nitroflurbiprofen, nitrofurantoin, nitrogen gas, nitroglycerin,nitroglycerine, nitroprusside, nitrous oxide, Nitrousoxide, nivocasan,nizatidine, N-Lite, N-monomethyl-L-arginine, Nobactine, nolatrexed,nolpitantium besilate, nomegestrol acetate, non-amlodipine calciumchannel blockers, nonoxynol-9, non-preserved latanoprost,norelgestromin, norepinephrine, norethandrolone, norethindrone,norethindrone acetate, norethisterone, norethisterone acetate,norethisterone enantate, norethrindone acetate, norfloxacin,norgestimate, nortriptylatine, nortriptyline, norursodeoxycholic acid,noscapine, novabupivacaine, Novasoy, Novotaks, NPC-18, NRTI, NS-2, NS-8,NTC-510A, NucleomaxX, nucleoside antiretroviral drugs, NVB, nystatin,Nystatin LF, Nyxol, O2, obatoclax, obeticholic acid, obicetrapib,obinepitide, OBT, ocaperidone, ocinaplon, octinoxate, octocrylene,octocrylene+tinosorb, octreotide acetate, odalasvir, odanacatib,odiparcil, Odyliresin, oestrogen, ofloxacin, oglemilast, oglufanidedisodium, olanzapine, olanzapine/fluoxetine, olaparib, olcegepant, oleicacid, olepra, olesoxime, oliceridine, OligoG CF-5/20, olmesartan,olmesartan cilexetil, olmesartan medoxomil, olodaterol, olodaterolhydrochloride, olopatadine, olopatadine hydrochloride, olprinone,olsalazine, oltipraz, omacetaxine mepesuccinate, omadacycline,omapatrilat, omarigliptin, omaveloxolone, ombitasvir, ombrabulin,omecamtiv mecarbil, omega, omega-3, omega-3 carboxylic acids, omega-3fatty acids, Omega-3 polyunsaturated fatty acids, omega-3-acid ethylesters, omega-3-carboxylic acids, omega-6, OmegaMAX, Omegaven,omeprazole, omeprazole and bicarbonate, omeprazole sodium,o-methylphenidate, omidenepag isopropyl, omigapil, Ommaya reservoir,Omri-Hep-B, onalespib, oncovin, ondansetron, ondelopran, opicapone,opipramol, opium, fumagillin, orantinib, orbofiban, Org-9426,orilotimod, oritavancin, orlistat, ornithine phenylacetate, orphenadrinecitrate, ortataxel, orteronel, Orthokine, orthosilicic acid, Orthostat,Orthostat-L, Orthovisc, orvepitant, oseltamivir, osemozotan, OSI-632,osilodrostat, ospemifene, OsteoDex, Osteonil, otenabant, oteracilpotassium, oteseconazole, otilonium bromide, oxacillin, oxaliplatin,oxandrolone, oxantel pamoate, oxazepam, oxcarbazepine, oxidizedglutathione sodium, Oximax, oxitriptan, oxitropium bromide, OX-NLA,oxybuprocaine, oxybutinin, oxybutynin, oxybutynin chloride, oxybutyninhydrochloride, oxycodone, oxycodone CR, oxycodone extended-release,oxycodone hydrochloride, oxycodone IR, Oxycyte, oxygen, oxymetazoline,oxymetazoline hydrochloride, oxymetholone, oxymorphone ER, oxymorphoneIR, oxypurinol, oxytocin, ozagrel, ozagrel hydrochloride, ozanimod,ozenoxacin, P-276-00, P-53, PAC-14028, paclitaxel, paclitaxelpoliglumex, paclitaxel-PM, pacritinib, pactimibe, pafuramidine,pagoclone, palanosetron hydrochloride, palbociclib, palifosfamide,paliperidone, paliperidone ER, paliperidone palmitate, paliroden,palivizumab, palonosetron, palovarotene, pamapimod, pamidronatedisodium, PAN-90806, Panavir 1, panobinostat, pantoprazole, pantothenicacid, pantovigar, papaverine, paquinimod, paracetamol, pardoprunox,parecoxib, paricalcitol, paritaprevir, parnaparin sodium, parogrelil,paromomycin, paroxetine, paroxetine hydrochloride, paroxetinehydrochloride hemihydrate, paroxetine mesylate, parthenolide, Passifloraincarnata, patidegib, patiromer calcium, patupilone, pazopanib,pazufloxacin, pazufloxacin mesylate, PCI-24781, PCI-27483, PD-110843,PD-115934, pectin, pefcalcitol, peficitinib, pegamotecan,pegcantratinib, pegylated liposomal doxorubicin, PEITC, pelitinib,pelitrexol, pelubiprofen, pemafibrate, pemetrexate, pemetrexed,pemetrexed disodium, pemirolast, pemirolast sodium, pemoline,penciclovir, penclomedine, penehyclidine hydrochloride, penicillamine,penicillin, penicillin G, penicillin V, pentaerythritol tetranitrate,PentaLyte, pentamidine, pentamidine isethionate, pentazocine,pentobarbital, pentosan polysulphate sodium, pentostatin, pentothal,pentoxifylline, pentoxyphilline, peramivir, perchlozone, peretinoin,perflubron emulsion, perflutren lipid microsphere, pergolide,perhexiline, perifosine, perillyl alcohol, perindopril, perindoprilarginine, permethrin, perospirone, perphenazine, perzinfotel, pethidine,pethidine hydrochloride, petrolatum, pexacerfont, pexidartinib,PF-04447943, PF-05089771, PF-05175157, PF-3654746, PF-3654764,PF-4191834, PF-4531083, PF-4691502, PF-489791, PF-610355, PG-2,PGL-2001, PGP/BCRP inhibitor, PH-797804, phenelzine, phenindione,pheniramine maleate, phenobarbital, phenobarbital sodium,phenoxybenzamine, phenprocoumon, phenserine, phentermine, phentolaminemesylate, phenylbutyrate, phenylephrine, phenylephrine hydrochloride,phenytoin, phloroglucinol, PHN-031, PHN-033, phosphate, phosphatidylserine, phosphatidylcholine, phosphatidylcholine-associated naproxen,phosphatidylcholine-encapsulated ibuprofen, Phospho-Lax, phosphorus,photopheresis, physostigmine, phytate, phytonadione, phytosterols,piboserod, pibrentasvir, picibanil, piclidenoson, piclozotan,picoplatin, picotamide, picroliv, picropodophyllin, picrorhiza,pictilisib, pilaralisib, pilocarpine, pilocarpine hydrochloride,pilsicainide, PIM-447, pimagedine, pimasertib hydrochloride,pimavanserin, pimecrolimus, pimodivir, pimozide, pindolol, pinocembrin,pioglitazone, pioglitazone hydrochloride, pipamperone, piperacillin,piperacillin sodium, piperacillin-tazobactam, piperaquine, piperaquinephosphate, piperine, piracetam, piragliatin, pirarubicin, pirenzepine,pirfenidone, piribedil, piridoxine, piritramide, piritrexim,piromelatine, piroxicam, pitavastatin, pitavastatin calcium, pitolisant,pivmeciellinam, pivmecillinam, pixantrone, PL-3994, plant sterols,platinum, plazomicin, pleconaril, plerixafor, plevitrexed, plinabulin,PLX-8394, PM-00104/50, PMI-001, PMK-N02RS1, pocapavir, polaprezinc,policosanol, polidocanol, polifeprosan 20 with carmustine, polmacoxib,polyethylene glycol, polyethylene glycolated IL-2, polyethyleneglycol-citrate-simethicone, polymeric nanoparticle docetaxel, polymyxinB sulphate, polyphenon E, polysaccharide-K, polysorbate 80, polysporin,polytetrafluoroethylene, pomaglumetad methionil, pomalidomide,ponatinib, ponesimod, poractant alpha, porfimer sodium, porfiromycin,posaconazole, posizolid, Posterisan akut, potassium, potassiumcanrenoate, potassium chloride, Potassium iodide, potassium nitrate,Potassium perchlorate, povidone, povidone iodine, pozanicline,poziotinib, PPA Lux 680, PPA-904, PPD-10558, PPI, PR-104, pracinostat,pradefovir, pradigastat, pralatrexate, pralidoxime, pralnacasan,pramiconazole, pramipexole, pranlukast, pranlukast hydrate, prasterone,prasugrel, pravastatin, praziquantel, prazosin, prednicarbate,prednisolone, prednisolone acetate, prednisolone phosphate, prednisolonesodium metazoate, prednisolone sodium succinate, prednisone, pregabalin,pregnenolone, preladenant, Premarin, Prempro, Prenatal vitamin,presatovir, pretomanid, prilocaine, primaquine, prinaberel, prinomastat,pritelivir, probenecid, Probiotics, probucol, procainamide, procaine,procaine hydrochloride, procarbazine, procarbazine hydrochloride,procaterol, procaterol hydrochloride, prochlorperazine, Procysteine,pro-docosapentaenoic acid, pro-eicosapentaenoic acid, progesterone,progestin, progestogen, progestogen dienogest, proglumide, proguanil,proguanil hydrochloride, Prolarix, promethazine, Prometra, Prometrium,Promisan, propacaine hydrochloride, propacetamol, propafenone,propafenone-SR, propanolol, proparacaine, propionyl-L-carnitine,propiverine, propiverine hydrochloride, propofol, Propofol Lipuro,propofol-lipuro, propranolol, propranolol hydrochloride, propranolol LA,propranolol XL, propylthiouracil, propyphenazone, Prosorba,prostaglandin, prostaglandin-E2, Prostin, Protelos, protriptyline,ProvideXtra, proxymetacaine, proxymetacaine hydrochloride, PRS-211375,prucalopride, prulifloxacin, Prurisol, pruvanserin, PRX-3140, PRX-8066,PSD-508, pseudoephedrine, pseudoephedrine hydrochloride, Pseudomonasaeruginosa mannose-sensitive hemagglutinin (PA-MSHA), PSI-5004, PSI-938,psyllium powder, PTH, p-toluene sulfonamide, puerarin, puerarin sodiumphosphate, pumosetrag, PVAC, PVP-ILH liposomes, PX-12, PYN-17,pyrantel-oxantel, pyrazinamide, pyridostigmine bromide, pyridoxal,pyridoxamine dihydrochloride, pyridoxine, pyridoxine hydrochloride,pyrimethamine, pyrimethamine % sulfadoxine, pyrimethamine/sulphadoxine,pyronaridine, Q-301, QAV-680, Qinbudan, Qizhitongluo, quercetin,quetiapine, quetiapine fumarate, quetiapine IR, quinacrine, quinagolidehydrochloride, quinapril hydrochloride, quinfamide, quinidine, quinine,quinolone, quinolones, quinupristin, quisinostat, quizartinibdihydrochloride, R-112, rabacfosadine, rabeprazole, rabeximod,rabusertib, racecadotril, raclopride, radafaxine, radalbuvir,radequinil, radezolid, Radha-108, radioactive iodine, Radioiodine,radiolabeled iodobenzamide, radiprodil, radium Ra 223 dichloride,radotinib, ragaglitazar, ralfinamide, raloxifene, raltegravir,raltitrexed, ramatroban, ramelteon, ramipril, ramosetron, ranibizumab,ranimustine, ranirestat, ranitidine, ranolazine, rapamycin, rasagiline,raseglurant, ravidasvir hydrochloride, ravuconazole, raxatrigine,razupenem, RBP-8000, RBx-10017609, RDEA-806, RDP-58, rebamipide,rebaudioside A, rebimastat, reboxetine, refametinib, reformulateddiclofenac, reformulated mebendazole, regadenoson, regorafenib,regrelor, relebactam, relenopride, relugolix, remifentanil, remimazolam,remimazolam tosylate, remogliflozin etabonate, REN-1654, renzapride,repaglinide, reparixin, repinotan, repurposed ajulemic acid, repurposedondansetron, resatorvid, reserpine, resiniferatoxin, resminostat,Resoferon, Respifor, Respimat, Restylane, Restylane SubQ, resveratrol,retagliptin, retapamulin, retaspimycin, retigabine, retinoic acid,retinoid, retinol, retosiban, retrovir, revamilast, revefenacin,revexepride, reviparin sodium, rezatomidine, RF-07026, rFSH, RG-4929,RG-7234, RG-7795, RGH-478, RGH-507, Rhenium-188-HEDP, Rhinox, RhuDex,ribavirin, ribociclib, riboflavin, ridaforolimus, ridinilazole,rifabutin, rifalazil, rifampicin, rifampin, rifamycin, rifapentine,rifaximin, rigosertib sodium, rikkunshito, rilapladib, rilmenidine,rilpivirine hydrochloride, riluzole, rimacalib, rimegepant, rimexolone,rimonabant, Ringer's acetate, Ringer's lactate solution, riociguat,ripasudil hydrochloride hydrate, risedronate sodium, risperidone,ritanserin, ritobegron ethyl ester hydrochloride, ritodrine, ritonavir,rituximab, rivaroxaban, rivastigmine, rivenprost, rivipansel sodium,rivoglitazone, rizatriptan, RLP-068, RNF43, RNS-60, RO-4929097,RO-5036505, robalzotan, rociletinib, rocuronium, rocuronium bromide,rofecoxib, Roferon-A, roflumilast, rolapitant, rolipram, rolofylline,ronacaleret, roniciclib, ronopterin, ropinirole, ropinirolehydrochloride, ropivacaine, roquinimex, rose bengal sodium,rosiglitazone, rosiglitazone maleate, rosiglitazone XR, rosiptoracetate, rostafuroxin, rostaporfin, rosuvastatin, rosuvastatin calcium,rotigotine, rovatirelin, roxadustat, roxithromycin, RP-323, RPh-201,RPL-554, RPM-02/08, RQ-00000004, R-salbutamol sulphate, rubitecan,ruboxistaurin, rucaparib, rucaparib camsylate, rucaparib phosphate,rufinamide, rupatadine, ruxolitinib, S Ketamine, S(+)-Ketamine, S-1,S-111, S-38093, S-707106, SAB-378, sabarubicin, Saccharomyces boulardii,sacubitril, S-adenosyl methionine, SAF-312, saffron, safinamide,safotibant, sagopilone, salbutamol, salbutamol HFA, salbutamol sulphate,salicylic acid, salicylic acid+benzoic acid, salidroside, saline,salirasib, salmon calcitonin, salsalate, Salubrin, samarium (153Sm)lexidronam, samatasvir, samidorphan, S-amlodipine gentisate,sapacitabine, sapanisertib, sapitinib, sapropterin, sapropterindihydrochloride, saquinavir, SAR-110894, saracatinib, sarecycline,saredutant, sargramostim, sarizotan hydrochloride, saroglitazar,sarpogrelate hydrochloride, satavaptan, satraplatin, saxagliptin,SB-773812, SBP-002, SC-49483, SCH-002063, SCH-497079, SCH-900776,schisandra sphenanthera extract, scopolamine, SCY-078, SDX-101,secnidazole, Sedlitzia rosmarinus, segesterone acetate, seladelparL-lysine dihydrate, selegiline, selegiline hydrochloride, selenium,selenium sulfide, selenomethionine, selepressin, seletracetam,selexipag, seliciclib, selinexor, selisistat, selodenoson, selonsertib,Selozok, selumetinib, selurampanel, semagacestat, semapimod, semaxanib,sembragiline, senicapoc, senna, Sensorcaine, Sentra PM, seocalcitol,sepantronium bromide, sepetaprost, sepranolone, septin, SER-150-DN,Serenoa repens, sergliflozin etabonate, serlopitant, serotonin reuptakeinhibitors, serotonin/norepinephrine reuptake inhibitors, sertaconazole,sertindole, sertraline, S-ethylisothiourea diethylphosphate, setileuton,setipiprant, S-etodolac, setrobuvir, sevelamer carbonate, sevelamerhydrochloride, sevoflurane, Sevofran, sevuparin sodium, SG-2000,Shanhuang Wuji decoction, SHP-465, sibrafiban, sibutramine, sildenafil,sildenafil citrate, Silexan, silibin, silibinin dihydrogensuccinate,silimarine, silodosin, silver nanoparticle, silver nitrate, silversulfadiazine, silybin, silymarin, simenepag isopropyl, simeprevir,simethicone, simeticone, simvastatin, Sinbaro, siponimod, sirolimus,sitafloxacin, sitagliptin, sitamaquine, sitaxentan, sitosterol,sivelestat, sivifene, s-ketamine, SKP-1052, SK-PC-B70M, SLx-4090,SMANCS, S-methionyl-L-citrulline, smilagenin, SMP-028, SN-38, SNX-5422,sodelglitazar, sodium, sodium 4-phenylbutyrate, sodium ascorbate, sodiumbenzoate, sodium bicarbonate, sodium butyrate, sodiumcarboxymethylcellulose, sodium chloride, sodium chromoglycate, sodiumcitrate, sodium dichloroacetate, sodium ferric gluconate complex, sodiumfluoride, sodium folinate, sodium fusidate, sodium hyaluronate, sodiumhydroxide, sodium hypochlorite, sodium ketorolac, sodium lactate, sodiumnitrate, sodium nitrite, sodium nitroprusside, sodium oxybate, sodiumphenylacetate, sodium phenylbutyrate, sodium phosphate, sodiumpicosulphate hydrate, sodium polystyrene sulfonate, sodium prasteronesulphate, sodium pyruvate, sodium stibogluconate, sodium sulfide, sodiumtetradecyl sulphate, sodium thiopental, sodium thiosulphate, sodiumvalproate, sodium-fluoride, sofinicline, sofosbuvir,sofosbuvir+daclatasvir, sofpironium bromide, solabegron, solcitinib,Soldesam, solifenacin, solithromycin, soluble ferric pyrophosphatecitrate, Solvazinc, somatostatin, sonedenoson, sonidegib, sonolisib,sorafenib, soraprazan, sorbitol, sorivudine, sotagliflozin, sotalol,sotrastaurin, sovaprevir, soy isoflavones, sparsentan, spebrutinib,spiramycin, spironolactone, sPLA2 inhibitors, SQ-109, squalamine,SR-T100, SSS, ST-101, STA-4783, standard bicarbonate, standardpneumoperitoneum, stannous compounds, stannsoporfin, statin, stavudine,stearidonic acid, S-tenatoprazole, Sterofundin, steroid, steroids,stiripentol, Stopain, Streptococcus faecium, Streptococcusthermophillus, streptomycin, streptozocin, strontium chloride Sr 89,strontium malonate, strontium ranelate, strontium-89, STW-5, STX-107,SU, SU-101, SU-14813, succimer, succinic acid, succinylcholine,sucralphate, sucralose, sucroferric oxyhydroxide, sucrose, Sufenta,sufentanil, sufentanil citrate, sufentanyl, sugammadex, sulbactam,sulbactam sodium, sulfadiazine, sulfadoxine, sulfadoxine+pyrimethamine,sulfalene-pyrimethamine, sulfamethoxazole, sulfasalazine, sulphate saltsolution, sulfonyl urea, sulfonylurea, sulfonylureas, sulforaphane,sulindac, sulodexide, sulopenem, sulopenem etzadroxil, sulphacetamidesodium, sulphadoxine, sulphadoxine-pyrimethamine, sulphamethoxazole,sulphonylurea, sulpiride, sultamicillin, sumanirole, sumatriptan,sumatriptan succinate, SUN-0597, SUN-1334H, sunitinib, suplatasttosilate, suramin, suramin sodium, surinabant, sutezolid, suxamethonium,SYI-2074, symbiotic, synbiotics, SYNSORB-Pk, synthetic hypericin, T/S,T-1225, T-2000, T3, tacalcitol, tacedinaline, tacrine, tacrolimus,Tacrolimus Hexal, tadalafil, tadekinig alpha, tafamidis, tafenoquine,tafluprost, tafoxiparin sodium, TAK-715, TAK-783, talabostat, taladegib,talampanel, talaporfin, talarozole, talazoparib, talc, TALL-104,talmapimod, talnetant, talniflumate, talotrexin, talsaclidine fumarate,taltirelin, tamibarotene, tamoxifen, tamsulosin, tamsulosinhydrochloride, tandospirone, tandutinib, tanespimycin, tanomastat,tanzisertib, tapentadol, taprenepag, Taradyal, tarafenacin, taranabant,tarenflurbil, taribavirin hydrochloride, tariquidar, tarloxotinibbromide, tasidotin HCl, tasimelteon, tasisulam, taspoglutide,tasquinimod, taurolidine, tauroursodeoxycholic acid, tavaborole,tavilermide, Taxol, Taxus, tazarotene, tazobactam, TC-2403, TC-3,TCM-606F, tebipenem pivoxil, tecadenoson, tecalcet, tecarfarin,tecastemizole, technetium bicisate, technetium etarfolatide, technetiumTc 99m tilmanocept, technetium Tc 99m trofolastat, technetium-99,tecovirimat, tedatioxetine, tedisamil, tedizolid phosphate, tegafur,tegaserod, teglarinad chloride, tegobuvir, teicoplanin, telaprevir,telapristone acetate, telatinib, telbivudine, telcagepant,telithromycin, telmisartan, telotristat etiprate, temazepam, temocapril,temocillin, temoporfin, temozolomide, temsirolimus, tenapanor,teneligliptin, teniposide, tenofovir, tenofovir alafenamide, tenofoviralafenamide fumarate, tenofovir disoproxil fumarate, tenofovir exalidex,tenofovir/emtricitabine, tenoxicam, teprenone, terameprocol, terazosin,terbinafine, terbinafine gel, terbinafine hydrochloride, terbogrel,terbutaline, terbutaline sulphate, terconazole, terguride, teriparatide,terlipressin acetate, terutroban, tesaglitazar, tesetaxel, tesevatinib,tesmilifene, tesofensine, Testagen, testosterone, testosteronecypionate, testosterone enanthate, testosterone undecanoate, Tetanustoxoid, tetomilast, tetrabenazine, tetracaine, tetracaine hydrochloride,tetracycline, tetracycline HCl, tetrahydrobiopterin,tetrahydrocannabinol, tetrathiomolybdate, Tetrodin, tezacaftor,tezacitabine, tezampanel, tezosentan, TG-100-115, TG-100801,thalidomide, THC, theophyllamine, theophylline, theophylline SR,theracurmin, thiamine, thiamine hydrochloride, thiazide, thiazidediuretics, thiazolidinedione, thiazolidinediones, thiocolchicoside,thioctic acid, thioguanine, thiopental, thiopental sodium, thiopentone,thioridazine, thiotepa, thiothixene, THR-0921, THR-4109, thrombin,thrombin microcapsules, thymoctonan, Thymoglobulin, thyroxine,tiagabine, tianeptine, tiapride, tibolone, ticagrelor, ticlopidine,tideglusib, tigecycline, tilapertin, tilarginine acetate, tiludronatedisodium, timapiprant, timcodar, timnodonic acid, timolol, timolol,timolol maleate, tinidazole, tinzaparin sodium, tiopronin, tiotropium,tiotropium bromide, tipelukast, tipifarnib, tipiracil hydrochloride,tipranavir, tirapazamine, tirasemtiv, tirilazad, tirofiban, tirofibanhydrochloride, titanium dioxide, TIVA, tivantinib, tivozanib,tizanidine, TMC-114/RTV, TMC-310911, TMC-647055, TNF-alpha, TNP-470,tobramycin, tocladesine, tocofersolan, tocopherol, tocopherols,tocopheryl phosphate mixture (TPM), tofacitinib, tofimilast,tofogliflozin, tolcapone, tolevamer, tolnaftato, tolperisone,tolterodine, tolterodine tartrate, tolvaptan, TOMM34, tonabersat,Tonalin SG1000T FFA, tonapofylline, Tongxinluo, topiramate,topiroxostat, topotecan, topotecan hydrochloride, torasemide,torcetrapib, toreforant, toremifene, tosedostat, tosufloxacin,tozadenant, tozasertib, trabectedin, trabectome, trabodenoson,tradipitant, tramadol, tramadol hydrochloride, tramazoline, trametinib,tramiprosate, trandolapril, tranexamic acid, tranilast,transcrocetinate-sodium, tranylcypromine, trastuzumab, travoprost,traxoprodil, trazadone, trazodone, trazodone hydrochloride, TRC-101,trehalose, trelagliptin succinate, trelanserin, treosulfan,treprostinil, treprostinil diolamine, tretinoin, triamcinolone,triamcinolone acetonide, triamcinolone hexacetonide, Triapine, TriatecHCT, Triaz, triazavirin, triazolam, tribendimidine, trichlormethiazide,trichlorothiazide, triciribine, tridolgosir, trientine, trientinehydrochloride, trifarotene, trifluoperazine, trifluridine, triflusal,triheptanoin, trihexyphenidyl, triiodothyronine, trilostane,trimebutine, trimebutine 3-thiocarbamoyl-benzenesulfonate, trimegestone,trimetazidine, trimethaphan, trimethobenzamide, trimethoprim,trimethoprim-sulfa, trimetrexate, trimipramine, trimipramine maleate,trinitrate, Triomune, Triplixam, tripotassium dicitrate bismuthate,tripterygium wilfordii, triptorelin acetate, triptorelin pamoate,trisodium citrate dihydrate, Trivax-AD, trofinetide, trofosfamide,troglitazone, tropicamide, tropisetron, trospium chloride,troxacitabine, troxipide, Trunature, trypsin-EDTA, TS-022, TTK, TTP-054,TTP-399, TTP-435, TTP-889, TTX-9401, tucaresol, tucidinostat,tulobuterol, TV-46763, TV-5010, TY-51924, tylenol, TZD, TZP-102,ubidecarenone, ubiquinol, ubrogepant, UCA-001, UCA-002, UCN-01,udenafil, UFT, UFT/LV, UFUR, UISH-001, UK-390957, UK-432097, UK-447841,ulimorelin, ulinastatin, ulipristal, ulobetasol, ulodesine,umeclidinium, umeclidinium bromide, umirolimus, unfractionated heparin,uniphyllin, upadacitinib, upamostat, uprifosbuvir, uprosertib, UR-906,uracil, urapidil, urea, uridine, uridine triacetate, URLC10,ursodeoxycholic acid, Ursolic acid, Urtica diociea, usistapide, UTD-1,utrogestan, V-116517, V-158866, V-404, VA-111913, vabicaserin,vaborbactam, vadadustat, vadimezan, Vagiprost, VAK-694, valaciclovir,valacyclovir, valbenazine, valdecoxib, valerian extract, valganciclovir,valium, valnivudine hydrochloride, valnoctamide stereoisomers,valomaciclovir stearate, valopicitabine dihydrochloride, valproate,valproic acid, valrocemide, valrubicin, valsartan, valsartan trisodiumhemipentahydrate, valspodar, valtorcitabine, vancomycin, vancomycinhydrochloride, vandetanib, vaniprevir, vanoxerine, vapendavir,vapitadine, vardenafil hydrochloride, varenicline, varespladib,varespladib methyl, varlitinib, vasopressin, vatalanib, vatiquinone,VDC-2008, vecuronium, vecuronium bromide, vedroprevir, VEGFR1, VEGFR1peptide, VEGFR1-A02-770, VEGFR2 peptide, VEGFR2-derived HLA-A0201,veliflapon, veliparib, velneperit, velpatasvir, velusetrag, vemurafenib,venetoclax, venlafaxine, venlafaxine hydrochloride, venlafaxine XR,vepoloxamer, verapamil, verapamil hydrochloride, vercirnon,verdiperstat, vericiguat, verinurad, vernakalant, vernakalanthydrochloride, verteporfin, verubulin, verucerfont, Verutex,vesatolimod, vesnarinone, vestipitant, vibegron, vicriviroc, Vidoca,vidofludimus, vidupiprant, vigabatrin, viitamin D, vilanterol,vilanterol trifenatate, vilaprisan, vilazodone, vildagliptin,vinblastin, vinblastine, vinblastine sulphate, vincristin, vincristine,vincristine sulphate, vindesine, vinflunine, vinorelbine, vinorelbineditartrate, vinpocetine, vintafolide, vipadenant, vismodegib,vistusertib, Vit B12, Vitamac, vitamin A, vitamin B, vitamin B1, vitaminB12, vitamin B-12, vitamin B2, vitamin B6, vitamin B-6, vitamin C,vitamin D, vitamin D2, vitamin D3, vitamin E, vitamin E succinatederivatives, vitamin K, vitamin K1, vitamins B1, vitamins B12, vitaminsB2, vitamins B6, Vitreosolve, Viusid, Vizomitin, vofopitant, voglibose,volasertib, volinanserin, vonoprazan fumarate, vorapaxar, voriconazole,vorinostat, vortioxetine hydrobromide, vosaroxin, voxilaprevir,voxtalisib, VP-101, VRx-3996, VSL-3, VVZ-149, VX-105, VX-135, VX-702,VX-710, Wafermine, warfarin, warfarin sodium, Water, WF-10, WH-1,Wobenzym, WX-554, xaliproden, xemilofiban, xenon, Xiang-sha-liu-jundecoction, xiaoqinglong, Xibrom, Xilei-San, ximelagatran, Xiyanping,XL-139, XP-21279, Xylitol, xylometazoline, Y-39983, Yallaferon,yonkenafil, yttrium clivatuzumab tetraxetan, yttrium Y 90 anti-CD66monoclonal antibody BW 250/183, zabofloxacin, zafirlukast, zalcitabine,zaleplon, zaltoprofen, zanamivir, ZD-6126, zeaxanthin, zibotentan,zicronapine, zidovudine, zileuton, zinc, zinc acetate, zinc oxide, zincpicolinate, zinc sulphate, zinc supplement, Zincas Forte, Zinthionein,ziprasidone, zofenopril, zoledronate, zoledronic acid, zoliflodacin,zolmitriptan, zolpidem, zolpidem tartrate, Zometa, zonampanel,zonisamide, zonisamide SR, zopiclone, zosuquidar, zotarolimus, zotepine,zucapsaicin, zuclopenthixol, and zuretinol acetate.

In some embodiments, the cargo of the engineered platelets descriedherein is a small molecule such as, but not limited to, those from theCeMM Library of Unique Drugs (CLOUD), as shown in Licciardello et al.,Nat Chem Biol; Vol. 13, pages 781-780 (2017)), which is herebyincorporated by reference in its entirety. For example, the smallmolecule may be, but it not limited to, Pinacidil, Altretamine,Pipobroman, Uracil Mustard, Trioxsalen, Plicamycin, Ambenonium,Edrophonium, Hexafluorenium, Oxtriphylline, Arbutamine, Guanabenz,Mephentermine, Methoxamine, Phenylpropanolamine, Protokylol,Tetrahydrozoline, Tolazoline, Bethanidine, Ergoloid, Oxprenolol,Penbutolol, Phentolamine, Propiomazine, Thiethylperazine, Fomepizole,Triamterene, Stanozolol, Dromostanolone, Ethylestrenol, Fluoxymesterone,Methyltestosterone, Deserpidine, Quinapril, Rescinnamine, Spirapril,Testolactone, Ethionamide, Sulfameter, Sulfacytine, Sulfamerazine,Sulfamethazine, Sulfamethizole, Sulfaphenazole, Sulfapyridine,Sulfathiazole, Sulfisoxazole, Sulfoxone, Cefmenoxime, Amdinocillin,Azlocillin, Bacampicillin, Carbenicillin, Cefalotin, Cefamandole,Cefditoren, Cefonicid, Ceforanide, Cefotiam, Cefpiramide, Cefradine,Ceftizoxime, Cephaloglycin, Cephapirin, Cyclacillin, Hetacillin,Loracarbef, Methicillin, Mezlocillin, Moxalactam, Nafcillin,Ticarcillin, Capreomycin, Demeclocycline, Dirithromycin, Methacycline,Oxytetracycline, Spectinomycin, Troleandomycin, Viomycin, Enoxacin,Novobiocin, Alatrofloxacin, Cinoxacin, Lomefloxacin, Nalidixic Acid,Sparfloxacin, Trovafloxacin, Acetohydroxamic Acid, Marinol,Ethoxzolamide, Acetohexamide, Fenoprofen, Oxyphenbutazone, Carprofen,Oxaprozin, Phenylbutazone, Tolmetin, Meclofenamic Acid,Methylergonovine, Acetophenazine, Carphenazine, Chlorprothixene,Mesoridazine, Triflupromazine, Promazine, Benzphetamine, Phenmetrazine,Chlorotrianisene, Estrone, Mestranol, Polyestradiol, Quinestrol,Cortisone, Fluprednisolone, Meprednisone, Paramethasone, Oxamniquine,Azatadine, Bromodiphenhydramine, Buclizine, Carbinoxamine,Chlophedianol, Dexbrompheniramine, Diphenylpyraline, Mepyramine,Methdilazine, Trimeprazine, Tripelennamine, Triprolidine, Romidepsin,Primidone, Butabarbital, Chlormezanone, Flurazepam, Glutethimide,Halazepam, Meprobamate, Metharbital, Methyprylon, Prazepam, Quazepam,Secobarbital, Talbutal, Thiamylal, Gamma Hydroxybutyric Acid, Memantine,Triclofos, Piperazine Hexahydrate, Desoxycorticosterone Pivalate,Pargyline, Carbachol, Oxyphenonium Bromide, Anisotropine, Clidinium,Cycrimine, Dicyclomine, Diphemanil, Ethopropazine, Fesoterodine,Hexocyclium, Isopropamide, Mepenzolate, Methantheline,Methylscopolamine, Metixene, Orphenadrine, Oxyphencyclimine,Procyclidine, Propantheline, Tridihexethyl, Trospium, Decamethonium,Pentolinium Tartrate, Cisatracurium Besylate, Succinylcholine Chloride,Doxacurium, Gallamine, Metocurine, Pancuronium, Pipecuronium,Rapacuronium, Tubocurarine, Guanadrel, Phendimetrazine, Anileridine,Difenoxin, Diphenoxylate, Levomethadyl, Oxymorphone, Propoxyphene,Levallorphan, Methylnaltrexone, (+/−)-Sulfinpyrazone, Pamidronic Acid,Risedronate, Tiludronate, Clofibrate, Dyphylline, Inamrinone,Vardenafil, Ethynodiol, Hydroxyprogesterone, Norethynodrel, UlipristalAcetate, Carboprost, Etretinate, Methysergide, Chlorphentermine,Acetyldigitoxin, Deslanoside, Chlorpropamide, Tolazamide, Tolbutamide,Methyclothiazide, Benzthiazide, Chlorothiazide, Cyclothiazide,Hydroflumethiazide, Polythiazide, Quinethazone, 5-Fluorouracil,Dextrothyroxine, Metyrosine, Rimantadine, Adefovir, Anisindione,Dicumarol, Nisoldipine, Trimethadione, Bepridil, Paramethadione,Bretylium Tosylate, Mephenytoin, Benzonatate, Ethotoin, Indecainide,Moricizine, Phenacemide, Tocainide, Pyrvinium Chloride Dihydrate,Halofantrine, Metaxalone, Diphenidol, Mebutamate, Chlorphenesin,Phensuximide, Thiabendazole, Benzquinamide, Piperacetazine,Ethchlorvynol, and Ethinamate.

IV. Production

In some embodiments, the engineered platelets described herein may beproduced using the technique described in Ito et al. (Cell, 174(3):636-648.e18, 2018, which is hereby incorporated by reference in itsentirety). Ito provides a method of clinical scale production ofplatelets from iPSC progenitors. Turbulence was observed to activateplatelet biogenesis for clinical scale ex vivo production of plateletsfrom human-induced pluripotent stem cells (iPSCs) (Ibid.). iPSCs derivedfrom immortalized megakaryocyte progenitor cell lines (imMKCLs) werecombined with soluble factors insulin Like Growth Factor Binding Protein2 (IGFBP2), macrophage migration inhibitory factor (MIF), and nardilysinconvertase (NRDC) in a bioreactor with control over the physicalparameters of turbulent energy and shear stress (Ibid.). Production ofgreater than 10¹¹ platelets were observed (Ibid.). Platelets wereobserved to function analogously to those derived from donors (Ibid.).

In certain embodiments of the invention herein, the imMKCL may beestablished by introducing cancer-derived MYC (c-MYC)/polycomb ringfinger proto-oncogene (BMI-1) and BCL2 1 like 1 (BCL-XL) genes into theiPSC using a lentivirus. Additional genes may be introduced or deletedresulting in an edited megakaryocyte, in fact even platelet specificpromoters have been previously characterized. These genes provideinducible gene expression in the presence of an agent, such asdoxorubicin (DOX). The imMKCL may be cyropreserved until cultivation isdesired. Megakaryocyte expansion is stimulated by contacting the cellline with the agent resulting expression of the inserted genes. Theagent is removed to halt gene expression and allow platelet production.

Current Federal Drug Administration (FDA)-approved rules for storage ofplatelets for transfusion require storage at 22° C. and must be usedwithin 6 days. Slichter et al. “Treatment of Bleeding in SeverelyThrombocytopenic Patients with Transfusion of Dimethyl Sulfoxide (DMSO)Cryopreserved Platelets (CPP) Is Safe—Report of a Phase 1 DoseEscalation Safety Trial”. Blood, 2016, which is hereby incorporated byreference in its entirety, hypothesizes cryopreservation is possible fortwo years when frozen with DMSO. After a positive phase 1 trial, phase 2and 3 trials are underway. Infusion of up to three sequential units ofcryopreserved platelets (CPP) in patients with severe thrombocytopeniaand active bleeding appeared to be “safe and without any evidence ofthrombotic complications despite CPP having a procoagulant phenotyperesulting from the cryopreservation process.” Therefore, cryopreservedplatelets likely have efficacy for stabilizing, reducing, or stoppingbleeding in thrombocytopenic patients as measured using the World HealthOrganization (WHO) bleeding grades. No evidence was found to underminethe hypothesis that cryopreserved platelets used for non-clottingpurposes would be as effective as platelets stored according to thepresent FDA rules.

V. Pharmaceutical Compositions

The present teachings further comprise pharmaceutical compositionscomprising one or more of the engineered platelets of the presentinvention, and optionally at least one pharmaceutically acceptableexcipient or inert ingredient. Further, a pharmaceutical may comprisethe therapeutic delivery system described herein.

As used herein the term “pharmaceutical composition” refers to apreparation of one or more of the engineered platelets described herein,or pharmaceutically acceptable salts thereof, optionally with otherchemical components such as physiologically suitable carriers andexcipients.

The term “excipient” or “inactive ingredient” refers to an inert orinactive substance added to a pharmaceutical composition to furtherfacilitate administration of a compound. Non-limiting examples of suchinert ingredients are disclosed herein under Formulations.

In some embodiments, compositions are administered to humans, humanpatients or subjects. For the purposes of the present disclosure, thephrase “active ingredient” generally refers to any one or more of theengineered platelets to be delivered as described herein.

Although the descriptions of pharmaceutical compositions provided hereinare principally directed to pharmaceutical compositions which aresuitable for administration to humans, it will be understood by theskilled artisan that such compositions are generally suitable foradministration to any other animal, e.g., to non-human animals, e.g.non-human mammals. Subjects to which administration of thepharmaceutical compositions is contemplated include, but are not limitedto, non-human mammals, including agricultural animals such as cattle,horses, chickens and pigs, domestic animals such as cats, dogs, orresearch animals such as mice, rats, rabbits, dogs and non-humanprimates.

A pharmaceutical composition in accordance with the invention may beprepared, packaged, and/or sold in bulk, as a single unit dose, and/oras a plurality of single unit doses. As used herein, a “unit dose” isdiscrete amount of the pharmaceutical composition comprising apredetermined amount of the active ingredient. The amount of the activeingredient is generally equal to the dosage of the active ingredientwhich would be administered to a subject and/or a convenient fraction ofsuch a dosage such as, for example, one-half or one-third of such adosage.

Relative amounts of the active ingredient, the pharmaceuticallyacceptable excipient or inert ingredient, and/or any additionalingredients in a pharmaceutical composition in accordance with theinvention will vary, depending upon the identity, size, and/or conditionof the subject treated and further depending upon the route by which thecomposition is to be administered. By way of example, the compositionmay comprise between 0.1% and 100%, e.g., between 0.5 and 50%, between1-30%, between 5-80%, at least 80% (w/w) active ingredient.

Efficacy of treatment or amelioration of disease can be assessed, forexample by measuring disease progression, disease remission, symptomseverity, reduction in pain, quality of life, dose of a medicationrequired to sustain a treatment effect, level of a disease marker or anyother measurable parameter appropriate for a given disease being treatedor targeted for prevention. It is well within the ability of one skilledin the art to monitor efficacy of treatment or prevention by measuringany one of such parameters, or any combination of parameters. Inconnection with the administration of compositions of the presentinvention, “effective against” for example a cancer, indicates thatadministration in a clinically appropriate manner results in abeneficial effect for at least a statistically significant fraction ofpatients, such as an improvement of symptoms, a cure, a reduction indisease load, reduction in tumor mass or cell numbers, extension oflife, improvement in quality of life, or other effect generallyrecognized as positive by medical doctors familiar with treating theparticular type of cancer.

A treatment or preventive effect is evident when there is astatistically significant improvement in one or more parameters ofdisease status, or by a failure to worsen or to develop symptoms wherethey would otherwise be anticipated. As an example, a favorable changeof at least 10% in a measurable parameter of disease, and preferably atleast 20%, 30%, 40%, 50% or more can be indicative of effectivetreatment. Efficacy for a given composition or formulation of thepresent invention can also be judged using an experimental animal modelfor the given disease as known in the art. When using an experimentalanimal model, efficacy of treatment is evidenced when a statisticallysignificant change is observed.

A. Therapeutic Delivery System

Various embodiments of the invention herein provide a non-thrombogenictherapeutic delivery system comprising at least one engineered platelet,also referred to as a SYNLET™ therapeutic delivery system.

SYNLET™ therapeutic delivery system may be produced using themegakaryocyte/platelet production strategies described herein.Progenitor cell can be sequentially edited with no consequences forsafety creating the possibility of designing entirely syntheticnetworks. Immunogenic cargo may be cloaked within the platelet followedby release at target site. Control antigen sensitivity throughdifferential loading of CPR-ITAMs and inhibitor CPR-ITIMs.

1. Treatment of Cancer

In some embodiments, the engineered platelets herein increasedspecificity may be used to treat solid tumors. In the field ofimmuno-oncology, the engineered platelets may be altered to be activatedby antigen specific T Cells to upregulate their function to clear tumorsexpressing defined neoantigens. Conversely, antigen specific T Cellsthat mediate autoimmune diseases could be targeted for destruction, withdefined antigens known in a variety of common diseases includingHashimoto's thyroiditis, type 1 diabetes and multiple sclerosis.

In one embodiment, the targeting moiety of the CPR may recognize CD19 todeliver chemotherapeutics locally. CD19 is a well-known B cell surfacemolecule, which upon B cell receptor activation enhances B-cell antigenreceptor induced signaling and expansion of B cell populations. CD19 isbroadly expressed in both normal and neoplastic B cells. Malignanciesderived from B cells such as chronic lymphocytic leukemia, acutelymphocytic leukemia and many non-Hodgkin lymphomas frequently retainCD19 expression. This near universal expression and specificity for asingle cell lineage has made CD19 an attractive target forimmunotherapies.

In some embodiments, the targeting moiety of a CPR may recognize a tumorspecific antigen (TSA), for example a cancer neoantigen that is onlyexpressed by tumor cells because of genetic mutations or alterations intranscription which alter protein coding sequences, therefore creatingnovel, foreign antigens. The genetic changes result from geneticsubstitution, insertion, deletion or any other genetic changes of anative cognate protein (i.e. a molecule that is expressed in normalcells).

For example, the tumor may be starved by the platelet causing clotformation to stop blood supply. Alternatively, CPR may target a tumor byexpression of an α-granule directed cargo or toxin mediating localdelivery of a therapeutic. In additional embodiments, the CPR may targeta tumor by expression of an α-granule directed antibody, such as acheckpoint inhibitor, to increase anti-tumor immunity.

The engineered platelets described herein may be engineered to killcancerous cells. For example, CD19 targeted TRAIL expressing plateletsthat treat cancerous B cell leukemias (BCL). CD19 targeted CAR-T cellshave shown great promise in the clinic versus BCL. TNF SuperfamilyMember (TRAIL) and Fas ligand (FASL) have been shown to induce BCL deathvia apoptosis upon CD40 stimulation (See, Dicker et al. “Fas-ligand(CD178) and TRAIL synergistically induce apoptosis of CD40-activatedchronic lymphocytic leukemia B cells”. Blood, 2005, which is herebyincorporated by reference in its entirety). CD40L is naturally exposedon activated platelets (see, Henn et al. “CD40 ligand on activatedplatelets triggers an inflammatory reaction of endothelial cells”.Nature, 1998, which is hereby incorporated by reference in its entirety)and could thus activate FASL/TRAIL dependent cell death pathways whenbound to BCL. FASL is naturally exposed on activated platelets (See,Schleicher et al. “Platelets induce apoptosis via membrane-bound FasL”.Blood, 2015, which is hereby incorporated by reference in its entirety).TRAIL expressing platelets have been used to decrease prostate cancermetastasis in mice (See, Li et al. “Genetic engineering of platelets toneutralize circulating tumor cells”. Journal of Controlled Release,2016, which is hereby incorporated by reference in its entirety). In oneembodiment, a resting platelet presenting a CD19-single-chain variablefragment (scFv)-ITAM and containing TRAIL, CD40L, and FASL ligands isactivated by binding of the CD19-scFv-ITAM with CD19 on a B cell.Activation results in the presentation of TRAIL, CD40L, and FASL on theplatelet surface. Platelet-induced death of leukemia cells is mediatedby binding of CD40L to the CD40 receptor of the B cell to activate theFASL/TRAIL-dependent cell death pathways.

In certain embodiments, platelets may be engineered to direct expansionof neoantigen specific T cells in vivo. Neoantigens are presented inmany human tumors and can be computationally identified. Expansion of Tcells ex vivo and reinfusion results in targeted tumor killing. Immunecheckpoint inhibition allows for T cells to kill tumors expressingneoantigens (however non-specificity results in severe side effects).Megakaryocytes can be loaded with MHC class 1 molecules with exogenouspeptides and transfer these to platelets. Neoantigens may be expressedin megakaryocytes, and an MHC class 1-ITAM fusion protein is able tostimulate checkpoint inhibitors. This would allow in vivo expansion ofneoantigen specific T cells. For example, a platelet may be engineeredto express MHC1-Neoantigen-ITAM. Both the engineered platelets and the Tcell are activated by interaction of the MHC1-Neoantigen-ITAM with aneoantigen specific T cell receptor (TCR). Activation results inpresentation of cytotoxic T-lymphocyte associated protein 4 (CTLA4) andprogrammed cell death 1 (PD-1) on the surface of the platelet andinteraction with CTLA4 inhibitor (CTLA4i) and PD-1 inhibitor (PD-li),respectively, on the T cell. Maximum T cell activation and expansion isreached by checkpoint blockade.

2. Treatment of Autoimmunity

In some embodiments, the engineered platelets described herein may beused to treat autoimmunity conditions. At least eighty-one autoimmunediseases have been identified in humans, and forty-five of theeighty-one disease have been associated with autoantigens withthirty-six of the autoantigens being tissue-specific. (See, Hayter, etal., Autoimmunity Reviews 11(2012) 754-765, which is hereby incorporatedby reference in its entirety). Autoimmune diseases were defined asdisorders where “1) the specific adaptive immune response is directed tothe affected organ or tissue; 2) autoreactive T cells and/orautoantibodies are present in the affected organ or tissue; 3)autoreactive T cells and/or autoantibodies can transfer the disease tohealthy individuals or animals; 4) immunization with the autoantigeninduces the disease in animal models: 5) elimination or suppression ofthe autoimmune response prevents disease progression or even amelioratesthe clinical manifestation.” (Ibid.) Additional criterion considered forthe definition was that antibody binding would disrupt functioning ofthe autoantigen (Ibid.). Self-tolerance check-points at each stage oflymphocyte development and activation have also been identified (Ibid.).

In some embodiments, the engineered platelets described herein may beused to suppress autoantigen specific T cells to treat autoimmunedisease. In some embodiments, CPRs in the engineered platelets mayinclude a region specific to a tissue associate with the autoantigen.For example, the tissue is selected from the group consisting of:adipose tissue, adrenal gland, ascites, bladder, blood, bone, bonemarrow, brain, cervix, connective tissue, ear, embryonic tissue,esophagus, eye, heart, intestine, kidney, larynx, liver, lung, lymph,lymph node, mammary gland, mouth, muscle, nerve, ovary, pancreas,parathyroid, pharynx, pituitary gland, placenta, prostate, salivarygland, skin, stomach, testis, thymus, thyroid, tonsil, trachea,umbilical cord, uterus, vascular, and spleen.

Table 12 shows the molecular target and/or tissue target for anon-exhaustive list of neurological system autoimmunity disorders fromHayter, et al.

TABLE 12 Neurological System Autoimmunity Disorders Disorder Tissueautoantibody Molecular target Autoimmune disseminated encephalomyelitisAnti-myelin MOG Autoimmune inner ear disease Anti-cochlear Battendisease/Neuronal Ceroid Lipofuscinoses Anti-GAD GAD65, GAD2 Chronicinflammatory demyelinating Anti-myelin Myelin associated glycoprotein,polyneuropathy MPZ Encephalitis lethargica Anti-sleptolysin-O Anti-basalganglia Guillain-Barr syndrome Anti-ganglioside GM1, Anti-PMP22 PMP22Hashimoto's Encephalopathy Anti-thyroid microsomal TPO/TG antibodies;anti-TPO Anti-TPO Isaac's syndrome/acquired neuromyotonia Anti-VGKC,Anti-AChR VGKC, KCNA6 Miller Fisher syndrome Anti-GQ1b; Anti-GM1;Anti-GD1a Morvan's syndrome Anti-VGKC; Anti-neuronal AChR KCNA1, VGKCMultiple sclerosis Anti-MOG; Anti-proteolipid MOG, MBP, NEFL Proteolipidprotein protein, myelin oligodendrocyte glycoprotein, myelin basicprotein, neurofilament light polypeptide. Myasthenia gravis Anti-AChR;MuSK CHRNA1; AChR Narcolepsy PANDAS Anti-TRIB2; Anti-neuronal; Anti-TRIB2 GlcNAc; Anti-Dnase B Rasmussen's encephalitis Anti-NMDA-type GluRNMDA-type GluR, GRIA3 Stiff-person syndrome Anti-GAD; Anti-amphiphysinGAD2 Vogt-Koyanagi-Harada syndrome Anti-Ku-Mel-1 KUMEL1, ARMC9

Table 13 shows the molecular target and/or tissue target for anon-exhaustive list of endocrine system autoimmunity disorders fromHayter, et al.

TABLE 13 Endocrine System Autoimmunity Disorders Disorder (system)Tissue autoantibody Molecular target Addison's diseaseAnti-21-hydroxylase; Anti-17 alpha-hydroxylase; CYP21A2, 21OHAnti-P450scc Autoimmune hypoparathyroidism Anti-CaSR Calcium sensingreceptor Autoimmune hypophysitis Anti-pituitary cytosolic proteinAutoimmune oophoritis Anti-OA; Anti-21OH Autoimmune orchitis Anti-NASPNuclear autoantigenic sperm protein Autoimmune polyglandularAnti-candidal enolase, anti-pituitary, anti-calcium syndrome I (APECED)sensing receptor protein, anti-aromatic L-amino acid decarboxylase,anti-tyrosine hydroxylase Autoimmune polyglandular Anti-21-hydroxylase;Anti-17 alpha-hydroxylase syndrome II Autoimmune polyglandularAnti-21-hydroxylase, anti-17 alpha-hydroxylase, syndrome IIIand-thyroperoxidase. Diabetes mellitus, type 1 Anti-GAD; Anti-insulin;Anti-ICA512; Anti-IA-2β Insulin, GAD65, PTPRN, Graves' disease TSIg;Anti-TBII TSHR, LMOD1 Hashimoto's autoimmune Anti-TPO; Anti-TGThyroperoxidase thyroiditis Immunodysregulation, polyendocrinopathy,enteropathy, X-linked

Table 14 shows the molecular target and/or tissue target for anon-exhaustive list of gastrointestinal system autoimmunity disordersfrom Hayter, et at.

TABLE 14 Gastrointestinal System Autoimmunity Disorders Disorder Tissueautoantibody Molecular target Autoimmune hepatitis Anti-smooth muscleantibody; SIMA, ASGPR type 1 Anti-nuclear antibody (ANA);(Asioglycoprotein receptor) Anti-actin Autoimmune hepatitis Anti-LKM-1;Anti-P-450 LKM1, CYP2D6 type 2 IID6 Autoimmune pancreatitisAnti-lactoferrin; Anti-amylase Lactoferrin alpha 2A; Anti-ACA-II Coeliacdisease Anti-TG2; Anti-gliadin Tissue trans-glutaminase 2 Crohn'sdisease ASCA Pernicious anemia/ Anti-H/K HK ATPase; ATP4A atrophicgastritis Primary biliary cirrhosis Anti-mitochondrial antibodies 74-kDaE2, KRT7, SP100, 74-kDa E2, keratin, sp100, actin. Primary sclerosingcholangitis pANCA Ulcerative colitis pANCA

Table 15 shows the molecular target and/or tissue target for anon-exhaustive list of hematopoietic autoimmunity disorders from Hayter,et al.

TABLE 15 Hematopoietic Autoimmunity Disorders Disorder Tissueautoantibody Molecular target Acquired hemophilia A Anti-FVIII FactorVIII Antiphospholipid syndrome Anti-cardiolipin; Beta2-GPI, Lupusanticoagulant; APOH Anti-b2GPI Autoinuatme hemolytic Anti-erythrocyteI/i Erythrocyte I/i, anemia RHCE/D Autoimmune Anti-erythrocyte; Anti-lymphoproliferative syndrome neutrophil Autoimmune neutropenia Anti-NA1;Anti-NA2 ITGB2; β2 Evans syndrome Anti-platelet; Anti- integrinerythrocyte Felty's syndrome Anti-G-CSF G-CSF Immune thrombocytopenicAnti-GpIIb/IIIa; Anti- ITGA2B, purpura ADAMTS13; Anti- ITGP3glycoprotein Ib-IX

Table 16 shows the molecular target and/or tissue target for anon-exhaustive list of musculoskeletal system autoimmunity disordersfrom Hayter, et al.

TABLE 16 Museuloskeletal System Autoimmunity Disorders Disorder Tissueautoantibody Molecular target Polymyositis/dermatomyositis Anti-Jo1;Anti-Mi-2; Mi-2, EXOSC10, HARS, Histidyl tRNA, aminoacyl tRNAAnti-CADM140 synthetase, DNA-dependent nucleosome-stimulated ATPase,EXOSC10 protein, chromodomain-helicase-DNA-binding protein 4 Relapsingpolychondritis Anti-collagen II; Anti- COL2A1 collagen IV; Anti-collagen IX Rheumatoid arthritis Rheumatoid factor; Fibrinogen, βα,PADI4, FGA Anti-CCP; Anti- collagen II Still's disease ANA;Anti-endothelial cell antibodies;

Table 17 shows the molecular target and/or tissue target for anon-exhaustive list of cutaneous and mucous autoimmunity disorders fromHayter, et al.

TABLE 17 Cutaneous and Mucous Autoimmunity Disorders Disorder Tissueautoantibody Molecular target Alopecia areata Anti-hair follicleTrichohyalin antibodies Bullous pemphigoid Anti-BP 180 BPAG1 (bullouspemphigoid associate glycoprotein 1) Cicatricial pemphigoid Anti-BP230Laminin-332, BPAG1 Dermatitis Anti-TGase3 TGM1 herpetiformis(transglutaminase) Discoid lupus erythematosus Epidermolysis bullosaAnti-type VII collagen; COL7A acquisita Anti-plectin Linear morpheaAnti-P80 Coilin P80 Coilin Pemphigus foliaceus Anti-Desmoglein IDesmoglein I Pemphigus vulgaris Anti-Desmoglein III Desmoglein IIIVitiligo Anti-MCHR1; SOX10 Anti-SOX-10

Table 18 shows the molecular target and/or tissue target for anon-exhaustive list of cutaneous autoimmunity disorders from Hayter, etal.

TABLE 18 Systemic Autoimmunity Disorders Disorder Tissue autoantibodyMolecular target Behçet disease Anti-oral mucous membrane Churg-Strausssyndrome cANCA Cogan's syndrome CREST syndrome Anti-centromereAnti-fibrillarin Essential mixed Anti-IgG; AECA cryoglobulinemia Mixedconnective tissue disease Anti-U1-ribonucleoprotein SNRNP70 POEMSsyndrome Anti-MAG; Anti-GM1 Scleroderma Anti-Scl70, anti-PM/Scl,anti-RNA TOP1, EXOSC10, TRIM21, SSB, polymerase III, anti-centromereTopoisomerase I, Ro, La, Ku, fibrillarin. Sjögren's syndrome ANA; SSB;SSA SSA/SSB, Ro, La, golgin Systemic lupus erythematosus Anti-dsDNA,Anti-U1A, Anti-U2B, SNRPB2, SNRPD1, PCNA, SNRPN, VIM, Anti-PCNA,Anti-Smith, Anti-SSA, TRIM21, SSB, U2 snRNP B, cardiolipin, Anti-SSB.fibronectin, Ro, La, histone H2A H2B, vimentin.

Table 19 shows the molecular target and/or tissue target for anon-exhaustive list of cardiovascular autoimmunity disorders fromHayter, et al.

TABLE 19 Cardiovascular Autoimmunity Disorders Disorder Tissueautoantibody Molecular target Erythema elevatum diutinum IgA ANCAKawasaki disease Microscopic polyangiitis pANCA MyeloperoxidasePolyarteritis nodosa Anti-endothelial cell antibodies Rheumatic feverAnti-streptolysin O; MYH6/7; cardiac Anti-DNase B myosin Takayasuarteritis Temporal arteritis Wegener's granulomatosis cANCA proteinase3, F2RL2

Table 20 shows the molecular target and/or tissue target for anon-exhaustive list of other autoimmunity disorders from Hayter, et al.

TABLE 20 Other Autoimmunity Disorders Disorder Tissue autoantibodyMolecular target HLA-B27-associated acute Anti-S-antigen SAG anterioruveitis Sympathetic ophthalmia Goodpasture's disease Anti-basementα3(IV)NC1 collagen, membrane GBM

In some embodiments, the engineered platelets comprise CPR with a regionrecognized by the autoreactive T cells that mediate the disease. Forexample, the CPR comprises an epitope from the molecular target inTables 12-20 loaded on to an MHC-ITAM fusion to directly target theautoreactive T cells. The engineered platelets may be loaded withcytotoxic or immunosuppressive protein or antibodies, which are releasedon activation of the platelet.

For instance, some cases of diabetes mellitus type 1 (T1DM) features Tcells specific to a particular insulin peptide. Therefore, using theMHC1-ITAM receptor fusion protein with an autoimmune driving peptide, ina platelet designed to release immunosuppressive factors would result inT cell specific immunosuppression. Exposure of an TL-2 receptor (IL-2R)to compete for IL-2, release of TGF-β1 or IL-10, and many otherpotential options on MHC1-ITAM activation mediates immunosuppressionsimilar to regulatory T (T_(reg)) cells.

In some embodiments, the engineered platelets comprising a CPR with amajor histocompatibility complex (MHC) class I or class H is used in thetreatment of an autoimmune disease. T cells expressing chimeric antigenreceptors (CAR) comprising the MHC ligand of a pathogenic T cellreceptor as an antigen binding domain of the CAR have been previouslyshown to be effective in the treatment of type 1 diabetes (T1D) (See,Perez et al., Immunology, 143, 609-617, which is hereby incorporated byreference in its entirety). In T1D, autoreactive CD8 and CD4 T cellsselectively destroy insulin-producing B cells in the pancreas (Ibid.).Some of the MHC-II-restricted epitopes recognized by the autoreactivecells have been observed to be derived from insulin/pre-proinsulin,islet-specific glucose-6-phosphatase catalytic subunit-related protein,glutamic acid decarboxylases 65 and 67, heat-shock proteins 60 and 70,insulinoma-associated protein 2, zinc transporter ZnT8, islet amyloidpolypeptide, chromogranin A, and other self antigens (Ibid.). Therefore,in some embodiments, the engineered platelets described herein include aCPR with a ligand or fragment thereof that will interact with theautoreactive cells to destroy the cells.

Some autoimmune conditions (e.g. autoimmune thyroiditis, pemphigusvulgaris) are driven by antibody dependent processes. CAR-T cells havebeen previously created to target the B cells that mediate autoimmunity(See, Ellebrecht et al. Science 2016). Engineered platelets describedherein expressing autoantigen-ITAM CPRs could kill B cells responsiblefor producing the autoantibodies driving autoimmunity. Activation of theengineered platelets on B cell binding could allow specific B cellkilling. An “AND” gate could be engineered to only permit activation ofthe engineered platelets in response to autoantigen B cell receptor(BCR) and CD19 binding, which would increase specificity of delivery ofcargo or toxin loaded into the platelet.

3. Treatment of Infection

Platelets have also been implicated in the clearance of bacterialinfections and constitute an important component of the innate immuneresponse, and thus SYNLET™ therapeutic delivery system could be used forthe treatment of drug resistant infections.

VI. Dosing and Administration

The present invention provides methods comprising administering any oneor more compositions for immunotherapy to a subject in need thereof.These may be administered to a subject using any amount and any route ofadministration effective for preventing or treating a clinical conditionsuch as cancer, infection diseases and other immunodeficient diseases.

Pharmaceutical compositions in accordance with the invention aretypically formulated in dosage unit form for ease of administration anduniformity of dosage. It will be understood, however, that the totaldaily usage of the compositions of the present invention may be decidedby the attending physician within the scope of sound medical judgment.The specific therapeutically effective, or prophylactically effectivedose level for any particular patient will depend upon a variety offactors including the disorder being treated and the severity of thedisorder; the activity of the specific compound employed; the specificcomposition employed; the age, body weight, general health, sex and dietof the patient; the time of administration, route of administration,previous or concurrent therapeutic interventions and rate of excretionof the specific compound employed; the duration of the treatment; drugsused in combination or coincidental with the specific compound employed;and like factors well known in the medical arts.

The term “effective amount” refers to the amount of the activeingredient needed to prevent or alleviate at least one or more signs orsymptoms of a specific disease and/or condition, and relates to asufficient amount of a composition to provide the desired effect. Theterm “therapeutically effective amount” therefore refers to an amount ofactive ingredient or a composition comprising the active ingredient thatis sufficient to promote a particular effect when administered to atypical subject. An effective amount would also include an amountsufficient to prevent or delay the development of a symptom of thedisease, alter the course of a symptom of the disease (for example butnot limited to, slow the progression of a symptom of the disease), orreverse a symptom of the disease. It is understood that for any givencase, an appropriate “effective amount” can be determined by one ofordinary skill in the art using routine experimentation.

The pharmaceutical, diagnostic, or prophylactic compositions of thepresent invention may be administered to a subject using any amount andany route of administration effective for preventing, treating,managing, or diagnosing diseases, disorders and/or conditions. The exactamount required will vary from subject to subject, depending on thespecies, age, and general condition of the subject, the severity of thedisease, the particular composition, its mode of administration, itsmode of activity, and the like. The subject may be a human, a mammal, oran animal. Compositions in accordance with the invention are typicallyformulated in unit dosage form for ease of administration and uniformityof dosage. It will be understood, however, that the total daily usage ofthe compositions of the present invention may be decided by theattending physician within the scope of sound medical judgment. Thespecific therapeutically effective, prophylactically effective, orappropriate diagnostic dose level for any particular individual willdepend upon a variety of factors including the disorder being treatedand the severity of the disorder; the activity of the specific payloademployed; the specific composition employed; the age, body weight,general health, sex and diet of the patient; the time of administration,and route of administration; the duration of the treatment; drugs usedin combination or coincidental with the active ingredient; and likefactors well known in the medical arts.

In certain embodiments, pharmaceutical compositions in accordance withthe present invention may be administered at dosage levels sufficient todeliver from about 0.01 mg/kg to about 100 mg/kg, from about 0.01 mg/kgto about 0.05 mg/kg, from about 0.05 mg/kg to about 0.5 mg/kg, fromabout 0.01 mg/kg to about 50 mg/kg, from about 0.1 mg/kg to about 40mg/kg, from about 0.5 mg/kg to about 30 mg/kg, from about 0.01 mg/kg toabout 10 mg/kg, from about 0.1 mg/kg to about 10 mg/kg, or from about 1mg/kg to about 25 mg/kg, of subject body weight per day, one or moretimes a day, to obtain the desired therapeutic, diagnostic, orprophylactic, effect.

The desired dosage of the pharmaceutical composition described hereinmay be delivered only once, three times a day, two times a day, once aday, every other day, every third day, every week, every two weeks,every three weeks, or every four weeks. In certain embodiments, thedesired dosage may be delivered using multiple administrations (e.g.,two, three, four, five, six, seven, eight, nine, ten, eleven, twelve,thirteen, fourteen, or more administrations). When multipleadministrations are employed, split dosing regimens such as thosedescribed herein may be used. As used herein, a “split dose” is thedivision of “single unit dose” or total daily dose into two or moredoses, e.g., two or more administrations of the “single unit dose”. Asused herein, a “single unit dose” is a dose of any therapeuticadministered in one dose/at one time/single route/single point ofcontact, i.e., single administration event.

In some embodiments, depending upon the nature of the engineeredplatelets, the engineered platelets may be introduced into a hostorganism, e.g., a mammal, in a wide variety of ways including byinjection, transfusion, infusion, local instillation or implantation. Insome aspects, the engineered platelets of the invention may beintroduced at the site of the tumor. The number of engineered plateletsthat are employed will depend upon a number of circumstances, thepurpose for the introduction, the lifetime of the engineered platelets,the protocol to be used, for example, the number of administrations, theability of the engineered platelets to multiply, or the like. Theengineered platelets may be in a physiologically-acceptable medium.

In some embodiments, the engineered platelets of the invention may beadministrated in multiple doses to subjects having a disease orcondition. The administrations generally effect an improvement in one ormore symptoms of cancer or a clinical condition and/or treat or preventcancer or clinical condition or symptom thereof.

The pharmaceutical compositions comprising the engineered platelets ofthe present invention may be administered by any route to achieve atherapeutically effective outcome.

These routes include, but are not limited to enteral (into theintestine), gastroenteral, epidural (into the dura matter), oral (by wayof the mouth), transdermal, peridural, intracerebral (into thecerebrum), intracerebroventricular (into the cerebral ventricles),epicutaneous (application onto the skin), intradermal, (into the skinitself), subcutaneous (under the skin), nasal administration (throughthe nose), intravenous (into a vein), intravenous bolus, intravenousdrip, intra-arterial (into an artery), intramuscular (into a muscle),intracranial (into the heart), intraosseous infusion (into the bonemarrow), intrathecal (into the spinal canal), intraperitoneal, (infusionor injection into the peritoneum), intrasinal infusion, intravitreal,(through the eye), intravenous injection (into a pathologic cavity)intracavitary (into the base of the penis), intravaginal administration,intrauterine, extra-amniotic administration, transdermal (diffusionthrough the intact skin for systemic distribution), transmucosal(diffusion through a mucous membrane), transvaginal, insufflation(snorting), sublingual, sublabial, enema, eye drops (onto theconjunctiva), in ear drops, auricular (in or by way of the ear), buccal(directed toward the cheek), conjunctival, cutaneous, dental (to a toothor teeth), electro-osmosis, endocervical, endosinusial, endotracheal,extracorporeal, hemodialysis, infiltration, interstitial,intra-abdominal, intra-amniotic, intra-articular, intrabiliary,intrabronchial, intrabursal, intracartilaginous (within a cartilage),intracaudal (within the cauda equine), intracisternal (within thecisterna magna cerebellomedularis), intracorneal (within the cornea),dental intracornal, intracoronary (within the coronary arteries),intracorporus cavernosum (within the dilatable spaces of the corporuscavernosa of the penis), intradiscal (within a disc), intraductal(within a duct of a gland), intraduodenal (within the duodenum),intradural (within or beneath the dura), intraepidermal (to theepidermis), intraesophageal (to the esophagus), intragastric (within thestomach), intragingival (within the gingivae), intraileal (within thedistal portion of the small intestine), intralesional (within orintroduced directly to a localized lesion), intraluminal (within a lumenof a tube), intralymphatic (within the lymph), intramedullary (withinthe marrow cavity of a bone), intrameningeal (within the meninges),intramyocardial (within the myocardium), intraocular (within the eye),intraovarian (within the ovary), intrapericardial (within thepericardium), intrapleural (within the pleura), intraprostatic (withinthe prostate gland), intrapulmonary (within the lungs or its bronchi),intrasinal (within the nasal or periorbital sinuses), intraspinal(within the vertebral column), intrasynovial (within the synovial cavityof a joint), intratendinous (within a tendon), intratesticular (withinthe testicle), intrathecal (within the cerebrospinal fluid at any levelof the cerebrospinal axis), intrathoracic (within the thorax),intratubular (within the tubules of an organ), intratumor (within atumor), intratympanic (within the aurus media), intravascular (within avessel or vessels), intraventricular (within a ventricle), iontophoresis(by means of electric current where ions of soluble salts migrate intothe tissues of the body), irrigation (to bathe or flush open wounds orbody cavities), laryngeal (directly upon the larynx), nasogastric(through the nose and into the stomach), occlusive dressing technique(topical route administration which is then covered by a dressing whichoccludes the area), ophthalmic (to the external eye), oropharyngeal(directly to the mouth and pharynx), parenteral, percutaneous,periarticular, peridural, perineural, periodontal, rectal, respiratory(within the respiratory tract by inhaling orally or nasally for local orsystemic effect), retrobulbar (behind the pons or behind the eyeball),intramyocardial (entering the myocardium), soft tissue, subarachnoid,subconjunctival, submucosal, topical, transplacental (through or acrossthe placenta), transtracheal (through the wall of the trachea),transtympanic (across or through the tympanic cavity), ureteral (to theureter), urethral (to the urethra), vaginal, caudal block, diagnostic,nerve block, biliary perfusion, cardiac perfusion, photopheresis orspinal.

VII. Definitions

At various places in the present specification, features or functions ofthe compositions of the present disclosure are disclosed in groups or inranges. It is specifically intended that the present disclosure includeeach and every individual sub combination of the members of such groupsand ranges. The following is a non-limiting list of term definitions.

As used herein, the term “antigen” is defined as a molecule thatprovokes an immune response when it is introduced into a subject orproduced by a subject such as tumor antigens which arise by the cancerdevelopment itself. This immune response may involve either antibodyproduction, or the activation of specific immunologically-competentcells such as cytotoxic T lymphocytes and T helper cells, or both.

As used herein, the term “approximately” or “about,” as applied to oneor more values of interest, refers to a value that is similar to astated reference value. In certain embodiments, the term “approximately”or “about” refers to a range of values that fall within 25, 20, 19, 18,17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or less ineither direction (greater than or less than) of the stated referencevalue unless otherwise stated or otherwise evident from the context(except where such number would exceed 100 of a possible value).

As used herein, the terms “associated with,” “conjugated,” “linked,”“attached,” and “tethered,” when used with respect to two or moremoieties, mean that the moieties are physically associated or connectedwith one another, either directly or via one or more additional moietiesthat serve as linking agents, to form a structure that is sufficientlystable so that the moieties remain physically associated under theconditions in which the structure is used, e.g., physiologicalconditions. An “association” need not be strictly through directcovalent chemical bonding. It may also suggest ionic or hydrogen bondingor a hybridization-based connectivity sufficiently stable such that the“associated” entities remain physically associated.

As used herein, the term “cancer” refers a broad group of variousdiseases characterized by the uncontrolled growth of abnormal cells inthe body. Unregulated cell division and growth results in the formationof malignant tumors that invade neighboring tissues ultimatelymetastasize to distant parts of the body through the lymphatic system orbloodstream.

As used herein, the term “cytokines” refers to a family of small solublefactors with pleiotropic functions that are produced by many cell typesthat can influence and regulate the function of the immune system.

As used herein, the term “delivery” refers to the act or manner ofdelivering a compound, substance, entity, moiety, cargo or payload. A“delivery agent” refers to any agent which facilitates, at least inpart, the in vivo delivery of one or more substances (including, but notlimited to a compound and/or compositions of the present invention) to acell, subject or other biological system cells.

As used herein, embodiments of the invention described herein are“engineered” when they are designed to have a feature or property,whether structural or chemical, that varies from a starting point, wildtype or native molecule.

As used herein, “expression” of a nucleic acid sequence refers to one ormore of the following events: (1) production of an RNA template from aDNA sequence (e.g., by transcription); (2) processing of an RNAtranscript (e.g., by splicing, editing, 5′ cap formation, and/or 3′ endprocessing); (3) translation of an RNA into a polypeptide or protein;(4) folding of a polypeptide or protein; and (5) post-translationalmodification of a polypeptide or protein.

As used herein, a “formulation” includes at least a compound and/orcomposition of the present invention and a delivery agent.

As used herein, a “fragment,” as used herein, refers to a portion. Forexample, fragments of proteins may comprise polypeptides obtained bydigesting full-length protein. In some embodiments, a fragment of aprotein includes at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15,16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85,90, 95, 100, 150, 200, 250 or more amino acids. In some embodiments,fragments of an antibody include portions of an antibody.

As used herein, the term “an immune cell” refers to any cell of theimmune system that originates from a hematopoietic stem cell in the bonemarrow, which gives rise to two major lineages, a myeloid progenitorcell (which give rise to myeloid cells such as monocytes, macrophages,dendritic cells, megakaryocytes and granulocytes) and a lymphoidprogenitor cell (which give rise to lymphoid cells such as T cells, Bcells and natural killer (NK) cells). Exemplary immune system cellsinclude a CD4+ T cell, a CD8+ T cell, a CD4− CD8− double negative Tcell, a T γδ cell, a Tap cell, a regulatory T cell, a natural killercell, and a dendritic cell. Macrophages and dendritic cells may bereferred to as “antigen presenting cells” or “APCs,” which arespecialized cells that can activate T cells when a majorhistocompatibility complex (MHC) receptor on the surface of the APCcomplexed with a peptide interacts with a TCR on the surface of a Tcell.

As used herein, the term “in vitro” refers to events that occur in anartificial environment, e.g., in a test tube or reaction vessel, in cellculture, in a Petri dish, etc., rather than within an organism (e.g.,animal, plant, or microbe).

As used herein, the term “in vivo” refers to events that occur within anorganism (e.g., animal, plant, or microbe or cell or tissue thereof).

As used herein, a “linker” or “targeting domain” refers to a portion ofa chimeric platelet receptor that recognizes and binds a desiredantigen.

As used herein, a “checkpoint factor” is any moiety or molecule whosefunction acts at the junction of a process. For example, a checkpointprotein, ligand or receptor may function to stall or accelerate the cellcycle.

As used herein, the term “messenger RNA” (mRNA) refers to anypolynucleotide which encodes a polypeptide of interest and which iscapable of being translated to produce the encoded polypeptide ofinterest in vitro, in vivo, in situ, or ex vivo.

As used herein, the term “mutation” refers to a change and/oralteration. In some embodiments, mutations may be changes and/oralterations to proteins (including peptides and polypeptides) and/ornucleic acids (including polynucleic acids). In some embodiments,mutations comprise changes and/or alterations to a protein and/ornucleic acid sequence. Such changes and/or alterations may comprise theaddition, substitution and or deletion of one or more amino acids (inthe case of proteins and/or peptides) and/or nucleotides (in the case ofnucleic acids and or polynucleic acids e.g., polynucleotides). In someembodiments, wherein mutations comprise the addition and/or substitutionof amino acids and/or nucleotides, such additions and/or substitutionsmay comprise 1 or more amino acid and/or nucleotide residues and mayinclude modified amino acids and/or nucleotides. The resultingconstruct, molecule or sequence of a mutation, change or alteration maybe referred to herein as a mutant.

As used herein, the term “neoantigen”, as used herein, refers to a tumorantigen that is present in tumor cells but not normal cells and do notinduce deletion of their cognate antigen specific T cells in thymus(i.e., central tolerance). These tumor neoantigens may provide a“foreign” signal, similar to pathogens, to induce an effective immuneresponse needed for cancer immunotherapy. A neoantigen may be restrictedto a specific tumor. A neoantigen be a peptide/protein with a missensemutation (missense neoantigen), or a new peptide with long, completelynovel stretches of amino acids from novel open reading frames (neoORFs).The neoORFs can be generated in some tumors by out-of-frame insertionsor deletions (due to defects in DNA mismatch repair causingmicrosatellite instability), gene-fusion, read-through mutations in stopcodons, or translation of improperly spliced RNA (e.g., Saeterdal etal., Proc Natl Acad Sci USA, 2001, 98: 13255-13260, which is herebyincorporated by reference in its entirety).

As used herein, the term “pharmaceutically acceptable excipient,” asused herein, refers to any ingredient other than active agents (e.g., asdescribed herein) present in pharmaceutical compositions and having theproperties of being substantially nontoxic and non-inflammatory insubjects. In some embodiments, pharmaceutically acceptable excipientsare vehicles capable of suspending and/or dissolving active agents.Excipients may include, for example: antiadherents, antioxidants,binders, coatings, compression aids, disintegrants, dyes (colors),emollients, emulsifiers, fillers (diluents), film formers or coatings,flavors, fragrances, glidants (flow enhancers), lubricants,preservatives, printing inks, sorbents, suspending or dispersing agents,sweeteners, and waters of hydration. Exemplary excipients include, butare not limited to: butylated hydroxytoluene (BHT), calcium carbonate,calcium phosphate (dibasic), calcium stearate, croscarmellose,crosslinked polyvinyl pyrrolidone, citric acid, crospovidone, cysteine,ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol,methionine, methylcellulose, methyl paraben, microcrystalline cellulose,polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinizedstarch, propyl paraben, retinyl palmitate, shellac, silicon dioxide,sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate,sorbitol, starch (corn), stearic acid, sucrose, talc, titanium dioxide,vitamin A, vitamin E, vitamin C, and xylitol.

Pharmaceutically acceptable salts of the compounds described herein areforms of the disclosed compounds wherein the acid or base moiety is inits salt form (e.g., as generated by reacting a free base group with asuitable organic acid). Examples of pharmaceutically acceptable saltsinclude, but are not limited to, mineral or organic acid salts of basicresidues such as amines; alkali or organic salts of acidic residues suchas carboxylic acids; and the like. Representative acid addition saltsinclude acetate, adipate, alginate, ascorbate, aspartate,benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate,camphorsulfonate, citrate, cyclopentanepropionate, digluconate,dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate,glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide,hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. Pharmaceutically acceptablesalts include the conventional non-toxic salts, for example, fromnon-toxic inorganic or organic acids. In some embodiments, apharmaceutically acceptable salt is prepared from a parent compoundwhich contains a basic or acidic moiety by conventional chemicalmethods. Generally, such salts can be prepared by reacting the free acidor base forms of these compounds with a stoichiometric amount of theappropriate base or acid in water or in an organic solvent, or in amixture of the two; generally, nonaqueous media like ether, ethylacetate, ethanol, isopropanol, or acetonitrile are preferred. Lists ofsuitable salts are found in Remington's Pharmaceutical Sciences, 17thed., Mack Publishing Company, Easton, Pa., 1985, p. 1418, PharmaceuticalSalts: Properties, Selection, and Use, P. H. Stahl and C. G. Wermuth(eds.), Wiley-VCH, 2008, and Berge et al., Journal of PharmaceuticalScience, 66, 1-19 (1977), each of which is incorporated herein byreference in its entirety.

As used herein, the term “subject” or “patient” refers to any organismto which a composition in accordance with the invention may beadministered, e.g., for experimental, diagnostic, prophylactic, and/ortherapeutic purposes. Typical subjects include animals (e.g., mammalssuch as mice, rats, rabbits, non-human primates, and humans) and/orplants.

As used herein, the term “T cell” refers to an immune cell that producesT cell receptors (TCRs).

As used herein, the term “T cell receptor” (TCR) refers to animmunoglobulin superfamily member having a variable antigen bindingdomain, a constant domain, a transmembrane region, and a shortcytoplasmic tail, which is capable of specifically binding to an antigenpeptide bound to a MHC receptor. A TCR can be found on the surface of acell or in soluble form and generally is comprised of a heterodimerhaving α and β chains (also known as TCRα and TCRβ, respectively), or γand δ chains (also known as TCRγ and TCRδ, respectively). Theextracellular portion of TCR chains (e.g., α-chain, β-chain) containstwo immunoglobulin domains, a variable domain (e.g., α-chain variabledomain or Vα, β-chain variable domain or Vβ) at the N-terminus, and oneconstant domain (e.g., α-chain constant domain or Cα and β-chainconstant domain or Cβ) adjacent to the cell membrane. Similar toimmunoglobulin, the variable domains contain complementary determiningregions (CDRs) separated by framework regions (FRs).

As used herein, the term “therapeutically effective amount” means anamount of an agent to be delivered (e.g., nucleic acid, drug,therapeutic agent, diagnostic agent, prophylactic agent, etc.) that issufficient, when administered to a subject suffering from or susceptibleto an infection, disease, disorder, and/or condition, to treat, improvesymptoms of, diagnose, prevent, and/or delay the onset of the infection,disease, disorder, and/or condition. In some embodiments, atherapeutically effective amount is provided in a single dose. In someembodiments, a therapeutically effective amount is administered in adosage regimen comprising a plurality of doses. Those skilled in the artwill appreciate that in some embodiments, a unit dosage form may beconsidered to comprise a therapeutically effective amount of aparticular agent or entity if it comprises an amount that is effectivewhen administered as part of such a dosage regimen.

As used herein, the terms “treatment” or “treating” denote an approachfor obtaining a beneficial or desired result including and preferably abeneficial or desired clinical result. Such beneficial or desiredclinical results include, but are not limited to, one or more of thefollowing: reducing the proliferation of (or destroying) cancerous cellsor other diseased, reducing metastasis of cancerous cells found incancers, shrinking the size of the tumor, decreasing symptoms resultingfrom the disease, increasing the quality of life of those suffering fromthe disease, decreasing the dose of other medications required to treatthe disease, delaying the progression of the disease, and/or prolongingsurvival of individuals.

As used herein, the term “therapeutic agent” refers to a biological,pharmaceutical, or chemical compound. Non-limiting examples includesimple or complex organic or inorganic molecule, a peptide, a protein,an oligonucleotide, an antibody, an antibody derivative, antibodyfragment, a receptor, and a soluble factor.

EQUIVALENTS AND SCOPE

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments in accordance with the invention described herein. The scopeof the present invention is not intended to be limited to the aboveDescription, but rather is as set forth in the appended claims.

In the claims, articles such as “a,” “an,” and “the” may mean one ormore than one unless indicated to the contrary or otherwise evident fromthe context. Claims or descriptions that include “or” between one ormore members of a group are considered satisfied if one, more than one,or all of the group members are present in, employed in, or otherwiserelevant to a given product or process unless indicated to the contraryor otherwise evident from the context. The invention includesembodiments in which exactly one member of the group is present in,employed in, or otherwise relevant to a given product or process. Theinvention includes embodiments in which more than one, or the entiregroup members are present in, employed in or otherwise relevant to agiven product or process.

It is also noted that the term “comprising” is intended to be open andpermits but does not require the inclusion of additional elements orsteps. When the term “comprising” is used herein, the term “consistingof” is thus also encompassed and disclosed.

Where ranges are given, endpoints are included. Furthermore, it is to beunderstood that unless otherwise indicated or otherwise evident from thecontext and understanding of one of ordinary skill in the art, valuesthat are expressed as ranges can assume any specific value or subrangewithin the stated ranges in different embodiments of the invention, tothe tenth of the unit of the lower limit of the range, unless thecontext clearly dictates otherwise.

In addition, it is to be understood that any particular embodiment ofthe present invention that falls within the prior art may be explicitlyexcluded from any one or more of the claims. Since such embodiments aredeemed to be known to one of ordinary skill in the art, they may beexcluded even if the exclusion is not set forth explicitly herein. Anyparticular embodiment of the compositions of the invention (e.g., anyantibiotic, therapeutic or active ingredient; any method of production;any method of use; etc.) can be excluded from any one or more claims,for any reason, whether or not related to the existence of prior art.

It is to be understood that the words which have been used are words ofdescription rather than limitation, and that changes may be made withinthe purview of the appended claims without departing from the true scopeand spirit of the invention in its broader aspects.

Described herein are compositions and methods for the design,production, administration, and/or formulation of engineered plateletsdescribed herein. In some embodiments, the engineered platelets maycarry cargo in the vesicles for delivery on activation by a target,which does not activate wild-type platelets. In some embodiments thenthe engineered platelets of the invention carny cargo in the vesiclesfor delivery on activation by a target, wherein the target does notactivate wild-type platelets. For example the target to which the CPRbinds is not a target that would typically activate wild-type platelets,but which does activate the engineered platelet through the interactionwith the target-binding CPR

The present invention is further illustrated by the followingnon-limiting examples. Although any materials and methods similar orequivalent to those described herein can be used in the practice ortesting of the present invention, the preferred materials and methodsare now described. Other features, objects and advantages of theinvention will be apparent from the description. In the description, thesingular forms also include the plural unless the context clearlydictates otherwise. Unless defined otherwise, all technical andscientific terms used herein have the same meaning as commonlyunderstood by one of ordinary skill in the art to which this inventionbelongs. In the case of conflict, the present description will control.

FIGURE LEGENDS

FIG. 1 —genome editing optimization/guide ID. A) Schematic of CRISPRguide selection and screening procedure. B) Guide KO generationefficiency (as predicted by Synthego ICE algorithm) within a pools ofiPSCs. C) Summary and repetition of highest efficiency guidenucleofection. N=2 per result, error bars indicate standard deviation.

FIG. 2 —Sequential editing process→7×KO. A) Schematic of sequentialknock-out approach. B) Quantification of viable cell number duringsequential KO approach. Viable cells identified based on exclusion of PIstain. C) Pooled knock-out efficiencies throughout sequential KOapproach. At each Cas9 RNP nucleofection event, half the cells weretaken for genomic DNA extraction and amplicons for all previous targetsites were amplified and screened for their KO level using Synthego ICE.

FIG. 3 —7×KO clone identification. A) Table showing Synthego ICE resultsfor gene KOs within clones produced from a single cell sort of the 7×KOpool. B) Repetition of Synthego ICE analysis on amplicons generated fromfurther expanded clones where results were absent in (A).

FIG. 4 —7×KO pool of cells forward programs towards a megakaryocyte likephenotype. A) Flow cytometry based MK differentiation marker panel andviability analysis on 7×KO pool 10 days post forward programminginduction using doxycycline. Performed in both unedited and 7×KO pool.B) As in (A), however 13 days post forward programming induction.

FIG. 5 —7×KO pool of cells is not activated by standard agonist. A)Microscopy images of unedited MKs and 7×KO pools, stained for P-Selectinat day 13 post doxycycline addition, following fixation. B) As in (A),however after the addition of TRAP6 (10 uM) and CRP (10 ug/ml) for 30minutes, followed by fixation. C) Flow cytometry assay of P-Selectinexposure in MKs stimulated with 300 ng/mL of PMA. Vehicle control or PMAwas added to live MKs and histograms shown are of P-Selectin staining7-10 minutes post agonist/vehicle addition. This assay was performed ona 7×KO clone (not pool) and done on day 15 post doxycycline addition.

FIG. 6 —Receptor design and lentiviral transduction. A) CPR receptordesign IDs. B) Schematic of CPR expression vectors packaged within inlentivirus. CPRs listed in A and mCherry expressed as a multicistronictranscript through the use of T2A sequence. Expression is driven by theEF1a promoter. C) Microscopy images of iPSCs transduced with lentivirusexpressing CPR sequences in (A), 2 days post transduction

FIG. 7 —Receptor expression on iPSC cell surface. A) CPR receptor designIDs. B) mCherry expression and CPR surface localisation as assayed byCD19-FITC based staining for CPR expression. 10 days post transductionwith lentivirus.

FIG. 8 —Receptor expressing cells FoP and retain expression. A) Flowcytometry based MK differentiation marker panel and viability analysison CPR3 expressing cells 10 and 16 days post forward programminginduction using doxycycline. B) CPR3 surface expression quantified usingFMC63-FITC staining of CPR3 expressing MKs and unstained MKs 10 dayspost doxycycline addition.

FIG. 9 —Receptor expressing MKs activate/degranulate in response toCD19+ve cells. A) Microscopy images of P-Selectin staining on fixed MKsexpressing CPR3 or untransduced controls following 30 minutes ofincubation with either BJABs (CD19+ve B cells) or Jurkats (CD19 negativeT cells). B) Flow cytometry quantification of P-Selectin staining ofsamples imaged in (A). C) MFI fold change of P-Selectin staining inindicated comparisons. MFI calculated following background subtraction,and performed within CD42 positive MK cell population.

FIG. 10 —schematic demonstrating the reduced thrombogenic potential ofthe platelets of the invention.

EXAMPLES Example 1. Establishing Platelet Production in a Laboratory

iPSC-iMKCL will be obtained from the Koji Eto Lab at MegakaryonCorporation (Kyoto office/Kyoto Lab: Kyoto Research Park, 93, Awatacho,Chudoji, Shimogyo-ku, Kyoto, 600-8815, JAPAN and the Tokyo office: 337Bldg #1, The University of Tokyo Institute of Medical Science 4-6-1,Shirokanedai, Minato-ku, Tokyo, 108-8639, JAPAN, in addition to aVERMES™ bioreactor (Satake Multimix) to allow rapid, high-qualityplatelet production.

Alternatively, a megakaryocyte line of choice, chosen after consultationwith key opinion leaders (KOLs) will be obtained and cultured. Back-upcell lines will be established and stored at −80° C. Platelet productionmay take place in a VERMES™ bioreactor, or in a shaking flask with thesix factors identified Ito et al., Cell, 174(3): 636-648.e18, 2018,which is hereby incorporated by reference in its entirety. The method ishypothesized to yield about 2.4×10⁶ platelets/ml in three days). Ahybrid approach combining the techniques described herein also may beused. For example, Meg01 cells (ATCC® CRL-2021™ from Sigma Aldrich) maybe combined with the six factors in a bioreactor with turbulence toresult in less clinical translation.

An in vitro assay for CD62 (specifically displayed on platelets onactivation) may be performed to ensure the platelets are active. Forexample, platelet CD62 will be measured using flow cytometry prior toactivation. Adenosine diphosphate (ADP), thrombin, or collagen will beadded to activate platelets, then percent of surface exposure of CD62will be measured.

Example 2. Generating Non-Thrombogenic Platelets

Once the progenitor cell line is established, it can be edited beforeplatelet production. Genes may be knocked out, such as genes that affectthe thrombogenicity of a platelet. Cas9 may be introduced to themegakaryocytes using a retrovirus to assist the editing process. Then,guide RNA (gRNA) electroporation would be performed. A tracking ofindels by decomposition (TIDE) analysis will be performed to confirm theknockout of desired regions.

The cloning efficiency of cells also will be measured to ensure thecells can be singly plated and grown up. In some embodiments of theinvention described herein, the function of the edited platelets will bemeasured using in vitro assays of platelet function, for example,microfluidic chips are commercially available to test aggregation.

Then, the platelets will be moved to in vivo function testing. A mousemodel as shown in Boulaftali et al. 2013, where endogenous mouseplatelets can be depleted, may be used (See, Boulaftali et al. “PlateletITAM signaling is critical for vascular integrity in inflammation”. JCI,2013, which is hereby incorporated by reference in its entirety). A lineof CLEC-2 knock-out (KO) human platelets will be generated to act as acontrol line.

The non-thrombogenic platelets (CLEC-2 and vascular endothelial cadherin(ve)) will be combined with a dye or beta-gal (β-Gal). Each mouse istransfused with a mix of control (CLEC-2) human platelets andnon-thrombogenic edited platelets. The mouse will be injured accordingto the protocol of an assay, such as hemoglobin (Hb) skin accumulationor tail vein bleeding time.

Any clot formed as a result of the assay will be observed for thepresence of edited platelets. The mice will be treated with rhodocytin(a snake venom component that acts through CLEC-2) to trigger CLEC-2dependent platelet aggregation of the edited platelets. Mice will beexamined for the presence of a clot. If no clot is present, the editedplatelets are truly non-thrombogenic.

Example 3. Generating CPR-Expressing Platelets

To test whether the edited platelets can be activated by an engineeredstimulus using a CPR, CPRs were designed between known ITAM containingplatelet receptors (GPVI, CLEC-2, and FCgR2A) and a model single chainantibody specific to an antigen (e.g. CD19). The construct will beintroduced either as an additional copy or by knock-in to the endogenousplatelet receptor locus to replace the cognate extracellular domain ofthe receptor. The CPR expressing platelets will be generated in vitroand exposed to a cell line expressing CD19 (e.g., NALM-6 cell line) anda control CD19 negative cell line (e.g., B16 melanoma cell line).

The ability of the CPR expressing platelets to subsequently activate inresponse to the presence of CD19 will be assayed in vitro throughmicroscopy. In some embodiments, a gene (e.g., TRAIL) will be expressedto increase cytotoxicity by the engineered platelets.

Using a similar technique, the CPR may be engineered to include portionsof known ITAM containing platelet receptors (GPVI, CLEC-2, and FCgR2A)and single chain MHC class 1 and MHC class 2 receptors. The variant ofMHC receptor used depends on the model used, e.g. New York esophagealsquamous cell carcinoma 1 (NY-ESO-1) from Astarte Biologics. Theconstruct may be introduced as either an additional copy or by knock-into the endogenous platelet receptor locus replacing its cognateextracellular domain. These CPR-expressing platelets will be produced invitro, and a peptide antigen will be added to the sample. TheCPR-expressing platelets will be exposed to a T-cell line responsive topeptide-MHC (or to a naïve batch of mixed T cells), and T cell responseto exposure will be observed. The platelets will be loaded withdifferent cytokine cocktails to determine whether the T cell responsecan be modified.

Example 5. Testing Non-Thrombogenic CPR-Expressing Platelets In Vivo

Non-thrombogenic platelets derived from a CD19 expressing melanoma cellline (or other melanoma cell line) will be engineered to contain CTLA4and PD-1 antibodies either passively or through retroviral transduction.Immunocompetent mice will be treated with these platelets and checkedfor melanoma treatment.

Using the CD19 Nalm-6 B Cell leukemia model, TRAIL will be expressed innon-thrombogenic platelets. FASL and CD40L are already present, whichsynergize with TRAIL to induce B Cell leukaemia death. NOD scid gammamice (NSG) mice having a tumor will be treated with the engineeredplatelets. The mice will be observed for a therapeutic benefit tovalidate the approach.

Alternatively, experimental autoimmune encephalomyelitis (EAE) may beinduced in mice using previously described protocol (vaccinated withmaltose binding protein (MBP)). Human platelets with mouse MHC and/orL8057 mouse cells with mouse MHC will be loaded with MBP peptide usedfor immunization. Further, platelets will be loaded with at least one ofcytotoxic components (to kill off specific cells) and TGF-β and otheranti-inflammatories. A well-defined clinical score system will be usedto establish whether the above is an effective model system for testingthe efficacy of non-thrombogenic CPR-expressing platelets in vivo.

Example 6—Materials and Methods for Example 7

CRISPR Guide Design

Guides were designed by identifying the first common exon of the targetexon of a gene. This exon was used as input to the CRISPOR algorithm forguide selection. Four guides per target gene were chosen based on theirdistribution across the exon and their specificity score, listed intable 21.Lentiviral iPSC TransductionReplication deficient lentiviral particles containing CPR constructs andmCherry were produced by Flash Therapeutics. hiPSC lines were routinelytransduced by 18-24 h single exposure to LVPs using multiplicity ofinfection of 100 in presence of 10 μg ml⁻¹ Protamine Sulfate (Sigma) inroutine culture medium.iPSC CloningHiPSCs were cloned by single cell sorting into 96 well plates. The dayprior to sorting, iPSCs were treated with CloneR (Stem CellTechnologies). 96 well plates were coated with Biolaminin 521 LN(Biolamina). CloneR was kept in the media until day 2 post sorting.Colonies were harvested 15-20 days post sorting, by treating wells withReLeSr and replating colonies into 24 well plates.Flow Cytometry and StainingSingle-cell suspensions were stained for 20 min at room temperatureusing combinations of FITC-, PE-, PE-Cy7-, APC-, and APC-H7-conjugatedantibodies. Background fluorescence were set againstfluorochrome-matched isotype control antibodies and compensationmatrices defined using single colour-stained cells.CRISPR Editing—Screening24 hours prior to nucleofection media was swapped for CloneR containingmedia. On the day of nucleofection, 1 μl of 61 pmol/μL of Alt-R HiFiCas9 V3 (Integrated DNA Technologies) was mixed with 2 μl of 91.5pmol/μL of sgRNA in TE (Synthego) (a 1:3 molar ratio) directly andincubated for at least 1 hour at room temperature. 100,000-500,000HiPSCs per nucleofection were harvested with GCDR (Stem CellTechnologies). Harvested cells were spun down and resuspended in 20 μLnucleofection buffer P3 (Lonza). Cas9/gRNA mix was then added to the 20μL cell/buffer P3 mix, then nucleofection was performed using 16-wellNucleocuvette Strip with 4D Nucleofector system (Lonza). Followingnucleofection, 80 μL of media was added to the nucleocuvette well, andcells were replated into a single well of a 24 well plate, in CloneRcontaining media. Media was changed two days later for mTeSR Plus.CRISPR Editing—Sequential24 hours prior to nucleofection media was swapped for CloneR containingmedia. On the day of nucleofection, 5 μl of 61 pmol/μL of Alt-R HiFiCas9 V3 (Integrated DNA Technologies) was mixed with 10 μl of 91.5pmol/μL of sgRNA in TE (Synthego) (a 1:3 molar ratio) directly andincubated for at least 1 hour at room temperature. 1-2.5 million HiPSCsper nucleofection were harvested with GCDR (Stem Cell Technologies).Harvested cells were spun down and resuspended in 100 μL nucleofectionbuffer P3 (Lonza). Cas9/gRNA mix was then added to the 100 μLcell/buffer P3 mix, then nucleofection was performed using the 100 μLNucleocuvette with 4D Nucleofector system (Lonza). Followingnucleofection, 400 μL of media was added to the nucleocuvette well, andcells were replated into two wells of a 6 well plate and one well of a24 well plate, in CloneR containing media. Media was changed two dayslater for mTeSR Plus. Cells were given 3-4 days total to recover, beforethe subsequent nucleofection was performed.CRISPR KO QuantificationGenotyping was performed by first harvesting HiPSC cells using GCDR orReLeSr. Genomic DNA was extracted using Kapa Express Extract Kit (Roche)following manufacturers instructions. Following genomic DNA extraction,the targeted genomic region was amplified using target locus specificprimers (See table 2). PCR fragments were PCR purified and submitted forSanger Sequencing (Source Bioscience). These sequences were then inputinto the ICE analysis software (Synthego) and thus editing efficiencieswere quantified.iPSC Cell Culture and Forward Programming to MKThe iPSC cell line RCIB-10 was forward programmed to megakaryocytes bythe concurrent expression of TAL1, FLI1 and GATA1 from a doxycyclineinducible promoter (see for example Dalby thesis, University ofCambridge “Forward programming of human pluripotent stem cells to amegakaryocyte-erythrocyte bi-potent progenitor population”; and Moreau14 Sep. 2017 “Forward Programming Megakaryocytes from Human PluripotentStem Cells” BBTS Annual Conference Glasgow 2017). The parental RCIB-10line was originally derived by episomal vector mediated expression ofhuman OCT4, SOX2, KLF4 and MYC reprogramming factors from the donor cellline.Cells were cultured under standard conditions with doxycycline for 10days at which point the cells were harvested.P-Selectin Based Activation Assay (CRP/TRAP-6/PMA)To assay the activation of MKs in response to mixing with knownagonists, 100,000-500,000 MKs were first harvested by centrifugation at100G for 8 minutes and resuspended in 100 μL of Tyrode's buffer (134 mMNaCl, 12 mM NaHCO3, 2.9 mM KCl, 0.34 mM Na2HPO4, 1 mM MgCl2, 10 mMHEPES, pH 7.4) containing anti P-Selectin antibody (Biolegend, cloneAK4, variable fluorophore at 1 μL/100 μL of cells). Where live cellswere assayed by flow, this was performed by direct sampling from thetube without resuspension of cells. Agonists were subsequently added andincubated with MKs for 40 minutes, before fixation with 1% PFA for 15minutes. Following PFA fixation, cells were resuspended in 300 μLTyrode's buffer containing anti-CD42 antibody (1 μL/100 μL) was added toallow for mature MK identification. MKs were analysed either by imagingusing confocal microscopy, or by flow cytometry. CRP (Cambcol) was addedto cells at a concentration of 10 μg/ml, TRAP-6 (Abcam) at aconcentration of 10 μM PMA (Sigma) at a concentration of 300 ng/mL. Whencells were used as agonists (Jurkats, DSMZ cat no; ACC 282 and BJABs—BCell lymphoma line, Ghevaert lab stock) they were added in 1:1 numbervs. Mks.

TABLE 21 gRNA primer sequences: HPS1 Exon 7 PAM Name Sequence sequencegrna1_HPS1_1r GGGGTGAATCAGTCGCTCCA GGG [SEQ ID NO: 56] grna2_HPS1_2GTCAACACCAGCCCCGAGCG GGG [SEQ ID NO: 57] grna3_HPS1_3GCTGGAGCGGCACGTCATCC AGG [SEQ ID NO: 58] grna4_HPS1_4rCTTGGAGTGCACGAGCAGGA AGG [SEQ ID NO: 59] ITGA2B Exon 7 PAM Name Sequencesequence grna5_ITGA2B_1r CAGTAGCCGTCGAAGTACTC TGG [SEQ ID NO: 60]grna6_ITGA2B_2 ATTTTCTCGAGTTACCGCCC AGG [SEQ ID NO: 61] grna7_ITGA2B_3rCTCGAGAAAATATCCGCAAC TGG [SEQ ID NO: 62] grna8_ITGA2B_3rGGGAGGACACGTGCCACAAA AGG [SEQ ID NO: 63] GP6 Exon 3 PAM Name Sequencesequence grna9_GP6_1 GGGCGTGGACCTGTACCGCC TGG [SEQ ID NO: 64]grna10_GP6_2r ACGAGCTCCAGCTGGTCGCT GGG [SEQ ID NO: 65] grna11_GP6_3rCGGAGGTCCCTGGCACCGGA GGG [SEQ ID NO: 66] grna12_GP6_4CCAGTGACCCTCCGGTGCCA GGG [SEQ ID NO: 67] Par1 Exon 2 PAM Name Sequencesequence grna13_Par1_1r GGAGCTGGTCAAATATCCGG AGG [SEQ ID NO: 68]grna14_Par1_2r TTCCTGAGAAGAAATGACCG GGG [SEQ ID NO: 69] grna15_Par1_3rACACTCCGGTGTACACAGAT GGG [SEQ ID NO: 70] grna16_Par1_4rACGATGGCCATGATGTTTAG TGG [SEQ ID NO: 71] Par4 Exon 2 PAM Name Sequencesequence grna17_Par4_1r ACTTGGCCTGGGTAGCCGCG GGG [SEQ ID NO: 72]grna18_Par4_2 GGTGCCCGCCCTCTATGGGC TGG [SEQ ID NO: 73] grna19_Par4_3TGGTGGGGCTGCCGGCCAAT GGG [SEQ ID NO: 74] grna20_Par4_4rAGCAGTGCCCGTGAGCTGTC CGG [SEQ ID NO: 75] Cox1 Exon 7 3′ and exon 8 PAMName Sequence sequence grna21_Cox1_1 ACTTCTGGCAAGATGGGTCC TGG[SEQ ID NO: 76] grna22_Cox1_2 TCACCAAGGCCTTGGGCCAT GGG [SEQ ID NO: 77]grna23_Cox1_3r TGTCTCCATAAATGTGGCCG AGG [SEQ ID NO: 78] grna24_Cox1_4AACTGCGGCTCTTTAAGGAT GGG [SEQ ID NO: 79] P2Y12 Exon 3 PAM Name Sequencesequence grna29_P2Y12_1r GTAGTCTCTGGTGCACAGAC TGG [SEQ ID NO: 80]grna30_P2Y12_2r GAAAGAAAATCCTCATCGCC AGG [SEQ ID NO: 81] grna31_P2Y12_3ATTCTTAGTGATGCCAAACT GGG [SEQ ID NO: 82] grna32_P2Y12_4rGATCGATAGTTATCAGTCCC AGG [SEQ ID NO: 83] B2M Exon 2 PAM Name Sequencesequence grna40_B2M_1r AAGTCAACTTCAATGTCGGA TGG [SEQ ID NO: 84]grna41_B2M_2r AGTCACATGGTTCACACGGC AGG [SEQ ID NO: 85] grna42_B2M_3ACTTGTCTTTCAGCAAGGAC TGG [SEQ ID NO: 86] grna43_B2M_4TCACGTCATCCAGCAGAGAA TGG [SEQ ID NO: 87]

TABLE 22 amplicon primers: HPS1 Sequencing primers RocO1_sHPS1_F1 FATCTGGTGCAGAGTCCAAGC [SEQ ID NO: 88] RocO1_sHPS1_R1 RTGGAGGAGGTGATTCTTGGC [SEQ ID NO: 89] Product 387 size: ITGA2BSequencing primers RocO3_ITGA2B_F1 F GGCTCCTGGCGGCTATTATT[SEQ ID NO: 90] RocO4_ITGA2B_R1 R CTTAGGCGGTGGGTTGGC [SEQ ID NO: 91]Product 360 size: ITGA2B Sequencing primers RocO5_GP6_F1 FAGCAGCGGGGTCCAGG [SEQ ID NO: 92] RocO6_GP6_R1 R CGTGGCACCACCACCC[SEQ ID NO: 93] Product 462 size: Par1 Sequencing primers RocO7_Par1_F1F ACCCACTCCTCCTAGTAAGAAAACA T [SEQ ID NO: 93] RocO8_Par1_R1 RCAAACTGCCAATCACTGCCG [SEQ ID NO: 94] Product 541 size: Par4Sequencing primers RocO9_Par4_F1 F ATGTCCAGCTGTTTCCCACC [SEQ ID NO: 95]RocO10_Par4_R1 R GCAGGTGGTAGGCGATCC [SEQ ID NO: 96] Product 415 size:Cox1 Sequencing primers RocO11_Cox1_F1 F CCAACCAGGGAAGAAGCAGT[SEQ ID NO: 97] RocO12_Cox1_R1 R TGGCACAAGCTTCCCACTC [SEQ ID NO: 98]Product 514 size: P2Y12 Sequencing primers RocO15_P2Y12_F1 FGAGGAGGCTGTGTCCAAAAA [SEQ ID NO: 99] RocO16_P2Y12_R1 RGGCTGCCTGTTGGTCAGAAT [SEQ ID NO: 100] Product 607 size: B2MSequencing primers RocO58_B2M_F1 F TGACACCAAGTTAGCCCCAA [SEQ ID NO: 101]RocO59_B2M_R1 R GGGATGGGACTCATTCAGGG [SEQ ID NO: 102] Product 463 size:

TABLE 23 (Media recipes): Mesoderm medium DMEM/F12, HEPES 500 ml(Thermofisher 7.5% NaHCO3 (Thermofisher) 3.6 ml 100x L-Ascorbic acid 5ml 2-phosphate sesquimagnesium salt hydrate (Sigma) Insulin-Transferrin-10 ml Selenium (ITS -G) 100x (but use as 50x) (Thermofisher) MK mediumIMDM without phenol 500 ml red 30% BSA (BioSera 8.4 ml SA-296)Insulin-Transferrin- 5 ml Selenium (ITS -G) 100x (Thermofisher) BetaMercaptoethanol 500 μl 55 mM

Example 7

To generate a non-thrombogenic, iPSC derived platelet-like chassis,genes encoding key components of the endogenous thrombotic process mustbe deleted. In this instance, the genes targeted were Cox1, GPVI, HPS1,ITGA2B, P2Y12, Par1 and Par4. CRISPR/Cas9 mediated IN/DEL generation waschosen as the method for gene knock-out (KO). First, guides weredesigned to target Cas9 nuclease to the above mentioned targets (FIG.1A). Four guides were designed per target, and nucleofected as complexwith the Cas9 protein into iPSCs, and their gene editing efficiencywithin the pool measured by Sanger sequencing and TIDE or the SynthegoICE algorithm. High efficiency guides resulting in >80% KO of eachtarget were identified in the guide screen (FIG. 1B). These guidesgenerated reproducibly high editing efficiency (FIG. 1C).To generate the non-thrombogenic chassis producing iPSC line, these KOsmust all be introduced into the same cell. To achieve this, a sequentialediting protocol was designed (FIG. 2A). In brief, Cas9 RNP complexesfeaturing the high efficiency guides identified previously werenucleofected into the same population of iPSCs sequentially, with 3-4days rest between each nucleofection. This protocol did not produce anadverse effect on cell viability or growth throughout the ˜3.5 weekprocess (FIG. 2B). Gene KO was quantified for each target hit previouslythroughout the sequential nucleofection protocol. No gene KO dilutionwas observed (as might occur if the KO itself was detrimental), andsurprisingly high gene editing efficiencies were observed for alltargets (>94% for all targets except COX1) (FIG. 2C). Following thesequential KO protocol, single cells were sorted into a 96 well plateand allowed to grow up forming clonal colonies. These colonies weresubsequently isolated and sequenced. Three 7×KO clones were identified(FIG. 3 ).Given the number of megakaryocyte (MK) specific genes KO'd within theseiPSCs, it remained unclear as to whether these iPSCs would still be ableto differentiate into MK like cells. To understand this, iPSCs wereforward programmed into MKs by doxycycline mediated induction of MKspecific transcription factors GATA1, TAL1 and FLI1. Cell surfaceexpression of known, well defined MK markers and viability was assayedduring the forward programming process (FIGS. 4A and B). This study wasperformed in the pool of 7×KO MKs, but given the exceedingly highediting efficiencies within the pool it is likely >90% of cells featureat least 6 KOs. We observed no effect on forward programming efficiencyor MK viability during the forward programming process. CD41 is ITGA2B,one of our target genes. Thus the lack of CD41 expression within the7×KO population validated the protein level KO of this gene as predictedby our sequencing based approach.To validate the non-thrombogenicity of our 7×KO MKs, and also theirretained function, we studied their degranulation response to knownplatelet agonists. MKs contain the same core signal transductionmachinery, plasma membranes and components as platelets (given plateletsare fragments of MKs), and thus MKs were used here as a surrogate foractual platelets. It is expected that the results seen in MKs wouldtranslate directly to platelets. To assay for degranulation, cellsurface P-Selectin exposure was used as a marker. P-Selectin is analpha-granule membrane protein, and is not usually present on theplatelet surface. Upon platelet activation, alpha-granules fuse with theplasma membrane and exocytose their contents (degranulation), and theirmembrane components mix with the plasma membrane. P-Selectin thusbecomes exposed and detectable by fluorescent antibody mediatedstaining. Resting 7×KO MKs feature lower basal levels of P-Selectinexposure than unedited wildtype MKs (FIG. 5A). Upon stimulation with twoclassical platelet agonists, CRP and TRAP6 (which signal through GPVIand PAR1 respectively—both KO'd in the 7×KO pool), no increase inP-Selectin staining was observed in the 7×KO MK pool. This is incontrast to the unedited MKs, which increased their P-Selectin and alsoappeared began to form small aggregates of cells (FIG. 5B). Importantly,upon stimulation of the 7×KO MKs with PMA, an agonist that bypasses thesignaling pathways removed within the 7×KO line, 7×KO MKs exposedP-Selectin as well if not better than unedited MKs (FIG. 5C). Takentogether, these activation experiments and the cell surface markerexperiments discussed previously demonstrate that deletion of ourcandidate non-thrombogenic genes in iPSCs does not perturb their abilityto differentiate into MK like cells, and does not disrupt the ability ofMKs to degranulate in response to non-deleted signal transductionmechanisms.Platelets contain ITAM domain containing receptors—specifically CLEC2,FCERG and FCGR2A. CLEC2 is a type-II membrane protein, whilst FCERG andFCGR2A are type-I membrane proteins. Type-I membrane proteins areamenable to fusion with scFV antibody domains (and other N-terminaltargeting mechanisms). Chimeric platelet receptors (CPRs) were thusdesigned as fusions between an scFV targeting the B cell antigen CD19derived from the FMC63 antibody, a hinge domain, and the transmembraneand cytoplasmic domains of FCERG and FCGR2A. This yielded four potentialreceptor designs (FIG. 6A). These designs were inserted into lentiviralexpression vectors as a multicistronic construct, with an mCherryfluorescent protein linked by a T2A peptide splitting sequence (FIG.6B). Viral particles were transduced onto iPSCs, and transductionefficiency examined by mCherry expression. Notable mCherry expressionwas detected across all four lentiviral expression vectors, and was notpresent in the untransduced control (FIG. 6C).To validate that the receptor itself was expressed and cell surfacelocalised, virally transduced iPSCs were stained with recombinant CD19fluorescently labelled with FITC. CD19-FITC should only label iPSCs ifthey express the anti-CD19 scFV on their cell surface, in the correctorientation. Notably, colonies positive for transduction (i.e. mCherrypositive) were also positive for CD19-FITC, indicating that the designedCPRs fold and correctly localise to the plasma membrane of the cellsexpressing them (FIG. 7 ).A clonal high CPR3 expressing iPSC line was forward programmed into MKs.Expression of the CPR3 construct did not impact the ability for iPSCs toforward program, as all classical MK specific markers were expressedwithin these cells. MK viability was not impacted by CPR3 expressioneither (FIG. 8A). Note that CD41 is clonally KO'd within these cells,and thus the lack of its expression is expected. To validate that CPR3was expressed and that this expression was maintained on the MK cellsurface, CD19-FITC staining was conducted (FIG. 8B). CPR surfaceexpression was observed, indicating MK differentiation did not silencethe lentiviral expression construct, or somehow alter receptorlocalisation.To study the functionality of the CPR, CPR3 expressing MKs and controluntransduced MKs were mixed with a CD19 expressing B cell leukaemia line(BJABs) or CD19 negative T cell leukaemia line (Jurkats) and P-Selectinexposure was measured as before. Microscopy imaging of mixed cellpopulations demonstrated increased P-Selectin exposure specificallywithin CPR3 expressing MKs when mixed with the CD19+ve BJABs (FIG. 9A).This was result was confirmed quantitatively by FACS based measurementof P-Selectin exposure (FIGS. 9B and C). BJAB cells do not activateuntransduced MKs, and CD19 negative Jurkats do not activate CPR3expressing MKs. These results demonstrate that the CPR3 constructspecifically stimulates MK degranulation in response to triggering byCD19 positive BJAB cells. Given that platelets are cytoplasmic fragmentsof MKs and the core signalling machinery is shared between them (giventhe shared cytoplasm), it is expected that these results shouldtranslate into platelets when produced from CPR3 expressing MKs.Additionally, given our observation that 7×KO MKs retain the ability toactivate and degranulate in response to agonists that have not had theircognate receptors deleted, it is expected that CPR3 expression within a7×KO line should trigger its degranulation upon mixing with CD19positive cells. Given the swappable nature of the external CPR targetingdomain, target specificity could be altered by swapping the anti-CD19scFV for alternative targeting mechanisms, while retaining the sameinternal signalling domain that has been shown here to trigger MKdegranulation on target engagement.Example Embodiments

1. A chimeric platelet receptor (CPR) comprising:

(a) a first region encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 1-19, 24-47, and 52-55; and

(b) a second region selected from the group consisting of: (i) a linkeror targeting domain encoded by a nucleic acid sequence selected from thegroup consisting of SEQ ID NO: 48-51; (ii) at least a portion of aprotein selected from the group consisting of: myelin oligodendrocyteglycoprotein (MOG), glutamic acid decarboxylase 2 (GAD65), myelinassociated glycoprotein (MAG), peripheral myelin protein 22 (PMP22),thyroid peroxidase (TPO), voltage-gated potassium channel (VGKC),proteolipid protein (PLP), acetylcholine receptor (AChR), tribblespseudokinase 2 (TRIB2), N-methyl-D-aspartate (NMDA)-type glutamatereceptor (GluR), glutamate decarboxylase 2 (GAD2), Armadillo repeatcontaining 9 (ARMC9), Cytochrome P450 Family 21 Subfamily A Member 2(CYP21A2), calcium sensing receptor (CASR), nuclear autoantigenic spermprotein (NASP), insulin, thyroid stimulating hormone receptor (TSHR),thyroperoxidase, asioglycoprotein receptor, Cytochrome P450 Family 2Subfamily D Member 6 (CYP2D6), lactoferrin (LF), tissuetrans-glutaminase (TTG), H/K ATP-ase, Factor XIII (F8),beta2-glycoprotein I (Beta2-GPI), erythrocyte I/I, B2 integrin (ITGB2),granulocyte-colony stimulating factor (G-CSF), glycoprotein (GP)IIb/IIa, collagen II (COLII), fibrinogen (FBG) βα, myeloperoxidase(MPO), cardiac myosin (CYO), proteinase 3 (PRTN3), trichohyalin (TCHH),bullous pemphigoid associated (BP), glycoprotein 1 (GPI), laminin-332(LM332), transglutaminase (TGM), type VII collagen (COLVII), P80 Coilin(COIL), Desmoglein I (DSG1), Desmoglein III (DSG3), SRY-Box 10 (SOX10),small nuclear ribonucleoprotein U1 subunit (70SNRNP70), S-antigen (SAG),and Collagen alpha-3(IV) chain (α3(IV)NC1 collagen); (iii) at least aportion of an antibody selected from the group consisting of; 3F8, 8H9,Abagovomab, Abciximab, Abituzumab, Abrezekimab, Abrilumab, Actoxumab,Adalimumab, Adecatumumab, Atidortoxumab, Aducanumab, Afasevikumab,Afelimomab, Alacizumab pego, Alemtuzumab, Alirocumab, Altumomabpentetate, Amatuximab, Anatumomab mafenatox, Andecaliximab, Anetumabravtansine, Anifrolumab, Anrukinzumab, Apolizumab, Aprutumab ixadotin,Arcitumomab, Ascrinvacumab, Aselizumab, Atezolizumab, Atinumab,Atorolimumab, Avelumab, Azintuxizumab vedotin, Bapineuzumab,Basiliximab, Bavituximab, BCD-100, Bectumomab, Begelomab, Belantamabmafodotin, Belimumab, Bemarituzumab, Benralizumab, Berlimatoxumab,Bermekimab, Bersanlimab, Bertilimumab, Besilesomab, Bevacizumab,Bezlotoxumab, Biciromab, Bimagrumab, Bimekizumab, Birtamimab,Bivatuzumab mertansine, Bleselumab, Blinatumomab, Blontuvetmab,Blosozumab, Bococizumab, Brazikumab, Brentuximab vedotin, Briakinumab,Brodalumab, Brolucizumab, Brontictuzumab, Burosumab, Cabiralizumab,Camidanlumab tesirine, Camrelizumab, Canakinumab, Cantuzumab mertansine,Cantuzumab ravtansine, Caplacizumab, Capromab pendetide, Carlumab,Carotuximab, Catumaxomab, cBR96-doxorubicin immunoconjugate,Cedelizumab, Cemiplimab, Cergutuzumab amunaleukin, Certolizumab pegol,Cetrelimab, Cetuximab, Cibisatamab, Cirmtuzumab, Citatuzumab bogatox,Cixutumumab, Clazakizumab, Clenoliximab, Clivatuzumab tetraxetan,Codrituzumab, Cofetuzumab pelidotin, Coltuximab ravtansine, Conatumumab,Concizumab, Cosfroviximab, Crenezumab, Crizanlizumab, Crotedumab,CR6261, Cusatuzumab, Dacetuzumab, Daclizumab, Dalotuzumab, Dapirolizumabpegol, Daratumumab, Dectrekumab, Demcizumab, Denintuzumab mafodotin,Denosumab, Depatuxizumab mafodotin, Derlotuximab biotin, Detumomab,Dezamizumab, Dinutuximab, Diridavumab, Domagrozumab, Dorlimomab aritox,Dostarlimab, Drozitumab, DS-8201, Duligotuzumab, Dupilumab, Durvalumab,Dusigitumab, Duvortuxizumab, Ecromeximab, Eculizumab, Edobacomab,Edrecolomab, Efalizumab, Efungumab, Eldelumab, Elezanumab, Elgemtumab,Elotuzumab, Elsilimomab, Emactuzumab, Emapalumab, Emibetuzumab,Emicizumab, Enapotamab vedotin, Enavatuzumab, Enfortumab vedotin,Enlimomab pegol, Enoblituzumab, Enokizumab, Enoticumab, Ensituximab,Epitumomab cituxetan, Epratuzumab, Eptinezumab, Erenumab, Erlizumab,Ertumaxomab, Etaracizumab, Etigilimab, Etrolizumab, Evinacumab,Evolocumab, Exbivirumab, Fanolesomab, Faralimomab, Faricimab,Farletuzumab, Fasinumab, FBTA05, Felvizumab, Fezakinumab, Fibatuzumab,Ficlatuzumab, Figitumumab, Firivumab, Flanvotumab, Fletikumab,Flotetuzumab, Fontolizumab, Foralumab, Foravirumab, Fremanezumab,Fresolimumab, Frovocimab, Frunevetmab, Fulranumab, Futuximab,Galcanezumab, Galiximab, Gancotamab, Ganitumab, Gantenerumab,Gatipotuzumab, Gavilimomab, Gedivumab, Gemtuzumab ozogamicin,Gevokizumab, Gilvetmab, Gimsilumab, Girentuximab, Glembatumumab vedotin,Golimumab, Gomiliximab, Gosuranemab, Guselkumab, Ianalumab, Ibalizumab,IBI308, Ibritumomab tiuxetan, Icrucumab, Idarucizumab, Ifabotuzumab,Igovomab, Iladatuzumab vedotin, IMAB362, Imalumab, Imaprelimab,Imciromab, Imgatuzumab, Inclacumab, Indatuximab ravtansine, Indusatumabvedotin, Inebilizumab, Infliximab, Intetumumab, Inolimomab, Inotuzumabozogamicin, Ipilimumab, Iomab-B, Iratumumab, Isatuximab, Iscalimab,Istiratumab, Itolizumab, Ixekizumab, Keliximab, Labetuzumab,Lacnotuzumab, Ladiratuzumab vedotin, Lampalizumab, Lanadelumab,Landogrozumab, Laprituximab emtansine, Larcaviximab, Lebrikizumab,Lemalesomab, Lendalizumab, Lenvervimab, Lenzilumab, Lerdelimumab,Leronlimab, Lesofavumab, Letolizumab, Lexatumumab, Libivirumab,Lifastuzumab vedotin, Ligelizumab, Loncastuximab tesirine, Losatuxizumabvedotin, Lilotomab satetraxetan, Lintuzumab, Lirilumab, Lodelcizumab,Lokivetmab, Lorvotuzumab mertansine, Lucatumumab, Lulizumab pegol,Lumiliximab, Lumretuzumab, Lupartumab amadotin, Lutikizumab,Mapatumumab, Margetuximab, Marstacimab, Maslimomab, Mavrilimumab,Matuzumab, Mepolizumab, Metelimumab, Milatuzumab, Minretumomab,Mirikizumab, Mirvetuximab soravtansine, Mitumomab, Modotuximab,Mogamulizumab, Monalizumab, Morolimumab, Mosunetuzumab, Motavizumab,Moxetumomab pasudotox, Muromonab-CD3, Nacolomab tafenatox, Namilumab,Naptumomab estafenatox, Naratuximab emtansine, Narnatumab, Natalizumab,Navicixizumab, Navivumab, Naxitamab, Nebacumab, Necitumumab,Nemolizumab, NEOD001, Nerelimomab, Nesvacumab, Netakimab, Nimotuzumab,Nirsevimab, Nivolumab, Nofetumomab merpentan, Obiltoxaximab,Obinutuzumab, Ocaratuzumab, Ocrelizumab, Odulimomab, Ofatumumab,Olaratumab, Oleclumab, Olendalizumab, Olokizumab, Omalizumab,Omburtamab, OMS721, Onartuzumab, Ontuxizumab, Onvatilimab, Opicinumab,Oportuzumab monatox, Oregovomab, Orticumab, Otelixizumab, Otilimab,Otlertuzumab, Oxelumab, Ozanezumab, Ozoralizumab, Pagibaximab,Palivizumab, Pamrevlumab, Panitumumab, Pankomab, Panobacumab,Parsatuzumab, Pascolizumab, Pasotuxizumab, Pateclizumab, Patritumab,PDR001, Pembrolizumab, Pemtumomab, Perakizumab, Pertuzumab, Pexelizumab,Pidilizumab, Pinatuzumab vedotin, Pintumomab, Placulumab, Plozalizumab,Pogalizumab, Polatuzumab vedotin, Ponezumab, Porgaviximab, Prasinezumab,Prezalizumab, Priliximab, Pritoxaximab, Pritumumab, PRO 140, Quilizumab,Racotumomab, Radretumab, Rafivirumab, Ralpancizumab, Ramucirumab,Ranevetmab, Ranibizumab, Raxibacumab, Ravagalimab, Ravulizumab,Refanezumab, Regavirumab, Relatlimab, Remtolumab, Reslizumab,Rilotumumab, Rinucumab, Risankizumab, Rituximab, Rivabazumab pegol,Robatumumab, Rmab, Roledumab, Romilkimab, Romosozumab, Rontalizumab,Rosmantuzumab, Rovalpituzumab tesirine, Rovelizumab, Rozanolixizumab,Ruplizumab, SA237, Sacituzumab govitecan, Samalizumab, Samrotamabvedotin, Sarilumab, Satralizumab, Satumomab pendetide, Secukinumab,Selicrelumab, Seribantumab, Setoxaximab, Setrusumab, Sevirumab,Sibrotuzumab, SGN-CD19A, SHP647, Sifalimumab, Siltuximab, Simtuzumab,Siplizumab, Sirtratumab vedotin, Sirukumab, Sofituzumab vedotin,Solanezumab, Solitomab, Sonepcizumab, Sontuzumab, Spartalizumab,Stamulumab, Sulesomab, Suptavumab, Sutimlimab, Suvizumab, Suvratoxumab,Tabalumab, Tacatuzumab tetraxetan, Tadocizumab, Talacotuzumab,Talizumab, Tamtuvetmab, Tanezumab, Taplitumomab paptox, Tarextumab,Tavolimab, Tefibazumab, Telimomab aritox, Telisotuzumab vedotin,Tenatumomab, Teneliximab, Teplizumab, Tepoditamab, Teprotumumab,Tesidolumab, Tetulomab, Tezepelumab, TGN1412, Tibulizumab,Tildrakizumab, Tigatuzumab, Timigutuzumab, Timolumab, Tiragotumab,Tislelizumab, Tisotumab vedotin, TNX-650, Tocilizumab, Tomuzotuximab,Toralizumab, Tosatoxumab, Tositumomab, Tovetumab, Tralokinumab,Trastuzumab, Trastuzumab emtansine, TRBS07, Tregalizumab, Tremelimumab,Trevogrumab, Tucotuzumab celmoleukin, Tuvirumab, Ublituximab,Ulocuplumab, Urelumab, Urtoxazumab, Ustekinumab, Utomilumab,Vadastuximab talirine, Vanalimab, Vandortuzumab vedotin, Vantictumab,Vanucizumab, Vapaliximab, Varisacumab, Varlilumab, Vatelizumab,Vedolizumab, Veltuzumab, Vepalimomab, Vesencumab, Visilizumab,Vobarilizumab, Volociximab, Vonlerolizumab, Vopratelimab, Vorsetuzumabmafodotin, Votumumab, Vunakizumab, Xentuzumab, XMAB-5574, Zalutumumab,Zanolimumab, Zatuximab, Zenocutuzumab, Ziralimumab, Zolbetuximab,Zolimomab aritox; and (iv) a major histocompatibility complex (MHC)class 1 receptor or a major histocompatibility complex (MHC) class 2receptor, wherein the MHC class 1 receptor is bound to a peptide derivedfrom a tumor antigen, a neoantigen, or an autoantigen or the MHC class 2receptor is bound to a peptide derived from a tumor antigen, aneoantigen, or an autoantigen.

2. The chimeric platelet receptor of embodiment 1, wherein the chimericplatelet receptor binds at least one antigen.

3. The chimeric platelet receptor of any one of embodiments 1 and 2,wherein the chimeric platelet receptor binds a tissue in the body of asubject.

4. The chimeric platelet receptor of any one of embodiments 1-3, whereinthe chimeric platelet receptor is an ITIM-containing receptor.

5. The chimeric platelet receptor of any one of embodiments 1-4, whereinthe chimeric platelet receptor is an ITAM-containing receptor.

6. A therapeutic delivery system comprising:

(a) an engineered platelet presenting the chimeric platelet receptor ofany of embodiments 1-5; and

(b) at least one therapeutic agent selected from the group consistingof; a toxin, a protein, a small molecule drug, and a nucleic acidpackaged within a vesicle inside the platelet.

7. The therapeutic delivery system of embodiment 6, wherein theengineered platelet is produced from an iPSC progenitor.

8. The therapeutic delivery system of any one of embodiments 6 and 7,wherein the nucleic acid is a mRNA, a miRNA, shRNA, and a clusteredregularly interspaced short palindromic repeats (CRISPR) sequence.

9. The therapeutic delivery system of any one of embodiments 6-8,wherein the protein is selected from the group consisting of anantibody, an enzyme, a cytokine, and a CRISPR associated protein 9(Cas9).

10. The therapeutic delivery system of embodiment 9, wherein the enzymeis a nuclease.

11. The therapeutic delivery system of embodiment 10, wherein thenuclease is a transcription activator-like effector nuclease (TALEN).

12. The therapeutic delivery system of embodiment 9, wherein theantibody binds a tumor antigen or a neoantigen.

13. The therapeutic delivery system of any one of embodiments 6-12,wherein the therapeutic agent is release from the platelet followingactivation of the platelet by an antigen recognized by the chimericplatelet receptor.

14. A method of treating a disease, disorder, or condition in a subject,the method comprising: administering to the subject the therapeuticdelivery system of any of embodiments 6-13, wherein the chimericreceptor is specific to an antigen associated with the disease,disorder, or condition.

15. The method of embodiment 14, wherein the disease, disorder, orcondition is selected from the group consisting of; a cancer, anautoimmunity, and an infection.

16. The method of any of embodiments 14 and 15, wherein the cancer isselected from the group consisting of: Acute granulocytic leukemia,Acute lymphocytic leukemia, Acute myelogenous leukemia, Adenocarcinoma,Adenosarcoma, Adrenal cancer, Adrenocortical carcinoma, Anal cancer,Anaplastic astrocytoma, Angiosarcoma, Appendix cancer, Astrocytoma,Basal cell carcinoma, B-Cell lymphoma), Bile duct cancer, Bladdercancer, Bone cancer, Bowel cancer, Brain cancer, Brain stem glioma,Brain tumor, Breast cancer, Carcinoid tumors, Cervical cancer,Cholangiocarcinoma, Chondrosarcoma, Chronic lymphocytic leukemia,Chronic myelogenous leukemia, Colon cancer, Colorectal cancer,Craniopharyngioma, Cutaneous lymphoma, Cutaneous melanoma, Diffuseastrocytoma, Ductal carcinoma in situ, Endometrial cancer, Ependymoma,Epithelioid sarcoma, Esophageal cancer, Ewing sarcoma, Extrahepatic bileduct cancer, Eye cancer, Fallopian tube cancer, Fibrosarcoma,Gallbladder cancer, Gastric cancer, Gastrointestinal cancer,Gastrointestinal carcinoid cancer, Gastrointestinal stromal tumors,General, Germ cell tumor, Glioblastoma multiforme, Glioma, Hairy cellleukemia, Head and neck cancer, Hemangioendothelioma, Hodgkin lymphoma,Hodgkin's disease, Hodgkin's lymphoma, Hypopharyngeal cancer,Infiltrating ductal carcinoma, Infiltrating lobular carcinoma,Inflammatory breast cancer, Intestinal Cancer, Intrahepatic bile ductcancer, Invasive/infiltrating breast cancer, Islet cell cancer, Jawcancer, Kaposi sarcoma, Kidney cancer, Laryngeal cancer, Leiomyosarcoma,Leptomeningeal metastases, Leukemia, Lip cancer, Liposarcoma, Livercancer, Lobular carcinoma in situ, Low-grade astrocytoma, Lung cancer,Lymph node cancer, Lymphoma, Male breast cancer, Medullary carcinoma,Medulloblastoma, Melanoma, Meningioma, Merkel cell carcinoma,Mesenchymal chondrosarcoma, Mesenchymous, Mesothelioma, Metastaticbreast cancer, Metastatic melanoma, Metastatic squamous neck cancer,Mixed gliomas, Mouth cancer, Mucinous carcinoma, Mucosal melanoma,Multiple myeloma, Nasal cavity cancer, Nasopharyngeal cancer, Neckcancer, Neuroblastoma, Neuroendocrine tumors, Non-Hodgkin lymphoma,Non-Hodgkin's lymphoma, Non-small cell lung cancer, Oat cell cancer,Ocular cancer, Ocular melanoma, Oligodendroglioma, Oral cancer, Oralcavity cancer, Oropharyngeal cancer, Osteogenic sarcoma, Osteosarcoma,Ovarian cancer, Ovarian epithelial cancer, Ovarian germ cell tumor,Ovarian primary peritoneal carcinoma, Ovarian sex cord stromal tumor,Paget's disease, Pancreatic cancer, Papillary carcinoma, Paranasal sinuscancer, Parathyroid cancer, Pelvic cancer, Penile cancer, Peripheralnerve cancer, Peritoneal cancer, Pharyngeal cancer, Pheochromocytoma,Pilocytic astrocytoma, Pineal region tumor, Pineoblastoma, Pituitarygland cancer, Primary central nervous system lymphoma, Prostate cancer,Rectal cancer, Renal cell cancer, Renal pelvis cancer, Rhabdomyosarcoma,Salivary gland cancer, Sarcoma, Sarcoma, bone, Sarcoma, soft tissue,Sarcoma, uterine, Sinus cancer, Skin cancer, Small cell lung cancer,Small intestine cancer, Soft tissue sarcoma, Spinal cancer, Spinalcolumn cancer, Spinal cord cancer, Spinal tumor, Squamous cellcarcinoma, Stomach cancer, Synovial sarcoma, T-cell lymphoma),Testicular cancer, Throat cancer, Thymoma/thymic carcinoma, Thyroidcancer, Tongue cancer, Tonsil cancer, Transitional cell cancer,Transitional cell cancer, Transitional cell cancer, Triple-negativebreast cancer, Tubal cancer, Tubular carcinoma, Ureteral cancer,Ureteral cancer, Urethral cancer, Uterine adenocarcinoma, Uterinecancer, Uterine sarcoma, Vaginal cancer, and Vulvar cancer.

17. The method of any of embodiments 14-16, further comprisingincubating the engineered platelet with the at least one therapeuticagent selected from the group consisting of: a toxin, a protein, and asmall molecule drug to produce the therapeutic delivery system.

18. The method of embodiment 17, wherein the nucleic acid is selectedfrom the group consisting of: a mRNA, a miRNA, shRNA, and a clusteredregularly interspaced short palindromic repeats (CRISPR) sequence.

19. The method of any one of embodiments 14-18, wherein the protein isselected from the group consisting of an antibody, an enzyme, and aCRISPR associated protein 9 (Cas9).

20. The method of embodiment 19, wherein the enzyme is a nuclease.

21. The method of embodiment 20, wherein the nuclease is a transcriptionactivator-like effector nuclease (TALEN).

22. The method of any of embodiments 17-21, wherein incubating occursprior to administering.

23. The method of any one of embodiments 14, 15, and 17-22, wherein thedisease, disorder, or condition is an autoimmunity selected from thegroup consisting of: Autoimmune disseminated encephalomyelitis,Autoimmune inner ear disease, Batten disease/Neuronal CeroidLipofuscinoses, Chronic inflammatory demyelinating polyneuropathy,Encephalitis lethargica, Anti-basal ganglia, Guillain-Barré syndrome,Hashimoto's Encephalopathy, Anti-TPO, Isaac's syndrome/acquiredneuromyotonia, Miller Fisher syndrome Morvan's syndrome, Multiplesclerosis, Myasthenia gravis, Narcolepsy PANDAS, Rasmussen'sencephalitis, Stiff-person syndrome, Vogt-Koyanagi-Harada syndrome,Addison's disease, Autoimmune hypoparathyroidism, Autoimmunehypophysitis, Autoimmune oophoritis, Autoimmune orchitis, Autoimmunepolyglandular syndrome I (APECED), Autoimmune polyglandular syndrome II,Autoimmune polyglandular syndrome III, Diabetes mellitus, type 1,Graves' disease, Hashimoto's autoimmune thyroiditis,Immunodysregulation, polyendocrinopathy, enteropathy, X-linked,Autoimmune hepatitis type 1, Autoimmune hepatitis type 2, Autoimmunepancreatitis, Coeliac disease, Crohn's disease, Perniciousanemia/atrophic gastritis, Primary biliary cirrhosis, Primary sclerosingcholangitis, Ulcerative colitis, Acquired hemophilia A, Antiphospholipidsyndrome, Autoimmune hemolytic anemia, Autoimmune lymphoproliferativesyndrome, Autoimmune neutropenia, Evans syndrome, Felty's syndrome,Immune thrombocytopenic purpura, Polymyositis/dermatomyositis, Relapsingpolychondritis, Rheumatoid arthritis, Still's disease, Alopecia areata,Bullous pemphigoid, Cicatricial pemphigoid, Dermatitis herpetiformis,Discoid lupus erythematosus, Epidermolysis bullosa acquisita, Linearmorphea, Pemphigus foliaceus, Pemphigus vulgaris, Vitiligo, Behçetdisease, Churg-Strauss syndrome, Cogan's syndrome, CREST syndrome,Anti-fibrillarin, Essential mixed cryoglobulinemia, Mixed connectivetissue disease, POEMS syndrome, Scleroderma, Sjögren's syndrome,Systemic lupus erythematosus, Erythema elevatum diutinum, Kawasakidisease, Microscopic polyangiitis, Polyarteritis nodosa, Rheumaticfever, Takayasu arteritis Temporal arteritis, Wegener's granulomatosis,HLA-B27-associated acute anterior uveitis, Sympathetic ophthalmia, andGoodpasture's disease.

24. An engineered platelet produced from a megakaryocyte comprising amutation in the nucleic acid sequence resulting in disruption of avesicle biogenesis pathway or a vesicle release pathway in the platelet,expression of a toxin, or deletion of a platelet receptor, mediator, orsignal transduction protein compared to a platelet produced from amegakaryocyte without the mutation.

25. The engineered platelet of embodiment 24, wherein the megakaryocyteis differentiated from an iPSC progenitor or the megakaryocyte isimmortalized.

26. The engineered platelet of any one of embodiments 24 and 25, whereinthe mutation occurs in a gene encoding a component of the vesiclebiogenesis pathway or a vesicle release pathway of the engineeredplatelet selected from the group consisting of: α-granules, densegranules, and large dense-core vesicle.

27. The engineered platelet of any one of embodiments 24-26, wherein thedeletion is of at least one gene selected from the group consisting of;Rab27a (RAS oncogene), HPS (haptoglobin) genes, integrin AIIbB3,GPIb-IX-V (Glycoprotein Ib complexed with glycoprotein IX), Par1(protease activated receptor 1), Par4 (protease activated receptor 4),P2Y 1 (purinergic receptor P2Y1), P2Y12 (purinergic receptor P2Y12), IP(PGI2R or prostaglandin 12 receptor), TP (TxA2R or Thromboxane A2Receptor), TLR (toll-like receptor), GPVI, a2B1 (type 1 collagenreceptor), GPIIbIIIA (Glycoprotein 11b Platelet Subunit Alpha), CLEC-2(C-type lectinlike receptor 2), MyD88 (Myeloid Differentiation PrimaryResponse 88), Galphaq (G-protein alpha pathway q), LIMK1 (LIM DomainKinase 1), vWF (von Willebrand), Fibrinogen, PDGF (platelet derivedgrowth factor), VEGF (vascular endothelial growth factor), Factor V,Factor VIII, Factor XI, Factor XIII, PF4 (platelet factor 4), NAP2(Nucleosome Assembly Protein 2), Prothrombin, High Molecular WeightKininogens, Plasminogen activator inhibitor 1, a2-antiplasmin,plasminogen, P-Selectin, CXCL4 (C-X-C motif chemokine ligand 4), CXCL7(C-X-C motif chemokine ligand 7), FGF (fibroblast growth factor), EGF(elongation growth factor), HGF (hepatocyte growth factor), IGF(insulin-like growth factor), Angipoetin, Thromboxane synthase, PAF(platelet activating factor), cPLA2a, Thromospondin, CD40L, SgIII(Secretogranin III), Endostatin, TGF-β (transforming growth factorbeta), Talin1, Kindlins, and ANO6 (Anoctamin 6).

28. The engineered platelet of any one of embodiments 24-27, wherein thedeletion is a knock-out of a gene encoding a pro-thrombotic factor.

29. The engineered platelet of embodiment 24, wherein the gene is a β2microglobulin gene, wherein the deletion results in endogenous MHC class1 disruption and the generation of a non-immunogenic platelet.

30. The engineered platelet of any one of embodiments 24-29, wherein themutation reduces the thrombogenic potential of the engineered plateletcompared to a platelet produced from a megakaryocyte without themutation.

31. A method of reducing activity in the immune system of a subject, themethod comprising:

(a) administering to the subject an engineered platelet presenting atleast one receptor expressing a major histocompatibility complex (MHC)molecule bound to a peptide derived from a tumor antigen, a neoantigen,or an autoantigen; and at least a portion of a domain from an ITAMreceptor.

32. The method of embodiment 31, wherein the receptor expresses an MHCclass I molecule.

33. The method of embodiment 31, wherein the receptor expresses an MHCclass II molecule.

34. The method of any one of embodiments 31-33, wherein the MHC moleculestimulates an immune response to an antigen.

35. The method of embodiment 34, wherein the antigen is associated withat least one disease, disorder, or condition selected from the groupconsisting of: a cancer, an autoimmunity, and an infection.

36. A method of in vitro production of platelets, the method comprising:

a) transfecting a plurality of induced pluripotent stem cell (iPSC)progenitors with an expression system, wherein the expression system isinduced by an agent not found in an iPSC;

b) establishing a megakaryocyte progenitor cell line by contacting theexpression system with the agent to expand megakaryocytes;

c) engineering the megakaryocyte to have at least one mutation selectedfrom the group consisting of: insertion of a nucleic sequence encoding achimeric platelet receptor of any one of embodiments 1-5, insertion of anucleic acid sequence encoding a toxin, and deletion of a nucleic acidsequence encoding a platelet receptor; and

d) removing the agent from the expression system to inducedifferentiation of the megakaryocytes into platelets.

37. The method of platelet production of embodiment 36, wherein themutation results in platelets with less immunogenicity compared toplatelets from human donors.

38. The method of platelet production of embodiment 37, wherein theplatelet does not function analogously to platelets derived from a humandonor.

39. The method of platelet production of any one of embodiments 36-38,wherein the deletion prevents release of cargo in the vesicles of theengineered platelets in response to endogenous platelet activationsignals.

40. The method of platelet production of any of embodiments 36-39,wherein the toxin is attached to an α-granule localization signal.

41. The method of platelet production of embodiment 40, wherein theα-granule localization signal directs the toxin to uptake into α-granulevesicles of the engineered platelet.

42. The method of platelet production of any one of embodiments 36-38,further comprising contacting the platelets with at least one selectedfrom the group consisting of: a toxin, and a small molecule drug underconditions to facilitate absorption by the platelet.

43. The method of platelet production of any one of embodiments 36-42,wherein the expression system further comprises a platelet-specificpromotors.

44. A method of in vivo gene editing or gene therapy in a subject, themethod comprising:

(a) administering to the subject an engineered platelet comprising achimeric platelet receptor of any one of embodiments 1-5 specific to atissue to be edited, wherein the engineered platelet is cloaking anadenovirus loaded with genome engineering machinery; and

(b) releasing the genome engineering machinery at the tissue.

45. The method of embodiment 44, wherein the genome engineeringmachinery is a CRISPR/Cas gene editing system.

46. A use of the therapeutic delivery system of any of embodiments 6-13,wherein the chimeric receptor is specific to an antigen associated withthe disease, disorder, or condition in treating a disease, disorder, orcondition in a subject.

47. The use of embodiment 46, wherein the disease, disorder, orcondition is selected from the group consisting of: a cancer, anautoimmunity, and an infection.

48. The use of embodiment 47, wherein the cancer is selected from thegroup consisting of: Acute granulocytic leukemia, Acute lymphocyticleukemia, Acute myelogenous leukemia, Adenocarcinoma, Adenosarcoma,Adrenal cancer, Adrenocortical carcinoma, Anal cancer, Anaplasticastrocytoma, Angiosarcoma, Appendix cancer, Astrocytoma, Basal cellcarcinoma, B-Cell lymphoma), Bile duct cancer, Bladder cancer, Bonecancer, Bowel cancer, Brain cancer, Brain stem glioma, Brain tumor,Breast cancer, Carcinoid tumors, Cervical cancer, Cholangiocarcinoma,Chondrosarcoma, Chronic lymphocytic leukemia, Chronic myelogenousleukemia, Colon cancer, Colorectal cancer, Craniopharyngioma, Cutaneouslymphoma, Cutaneous melanoma, Diffuse astrocytoma, Ductal carcinoma insitu, Endometrial cancer, Ependymoma, Epithelioid sarcoma, Esophagealcancer, Ewing sarcoma, Extrahepatic bile duct cancer, Eye cancer,Fallopian tube cancer, Fibrosarcoma, Gallbladder cancer, Gastric cancer,Gastrointestinal cancer, Gastrointestinal carcinoid cancer,Gastrointestinal stromal tumors, General, Germ cell tumor, Glioblastomamultiforme, Glioma, Hairy cell leukemia, Head and neck cancer,Hemangioendothelioma, Hodgkin lymphoma, Hodgkin's disease, Hodgkin'slymphoma, Hypopharyngeal cancer, Infiltrating ductal carcinoma,Infiltrating lobular carcinoma, Inflammatory breast cancer, IntestinalCancer, Intrahepatic bile duct cancer, Invasive/infiltrating breastcancer, Islet cell cancer, Jaw cancer, Kaposi sarcoma, Kidney cancer,Laryngeal cancer, Leiomyosarcoma, Leptomeningeal metastases, Leukemia,Lip cancer, Liposarcoma, Liver cancer, Lobular carcinoma in situ,Low-grade astrocytoma, Lung cancer, Lymph node cancer, Lymphoma, Malebreast cancer, Medullary carcinoma, Medulloblastoma, Melanoma,Meningioma, Merkel cell carcinoma, Mesenchymal chondrosarcoma,Mesenchymous, Mesothelioma, Metastatic breast cancer, Metastaticmelanoma, Metastatic squamous neck cancer, Mixed gliomas, Mouth cancer,Mucinous carcinoma, Mucosal melanoma, Multiple myeloma, Nasal cavitycancer, Nasopharyngeal cancer, Neck cancer, Neuroblastoma,Neuroendocrine tumors, Non-Hodgkin lymphoma, Non-Hodgkin's lymphoma,Non-small cell lung cancer, Oat cell cancer, Ocular cancer, Ocularmelanoma, Oligodendroglioma, Oral cancer, Oral cavity cancer,Oropharyngeal cancer, Osteogenic sarcoma, Osteosarcoma, Ovarian cancer,Ovarian epithelial cancer, Ovarian germ cell tumor, Ovarian primaryperitoneal carcinoma, Ovarian sex cord stromal tumor, Paget's disease,Pancreatic cancer, Papillary carcinoma, Paranasal sinus cancer,Parathyroid cancer, Pelvic cancer, Penile cancer, Peripheral nervecancer, Peritoneal cancer, Pharyngeal cancer, Pheochromocytoma,Pilocytic astrocytoma, Pineal region tumor, Pineoblastoma, Pituitarygland cancer, Primary central nervous system lymphoma, Prostate cancer,Rectal cancer, Renal cell cancer, Renal pelvis cancer, Rhabdomyosarcoma,Salivary gland cancer, Sarcoma, Sarcoma, bone, Sarcoma, soft tissue,Sarcoma, uterine, Sinus cancer, Skin cancer, Small cell lung cancer,Small intestine cancer, Soft tissue sarcoma, Spinal cancer, Spinalcolumn cancer, Spinal cord cancer, Spinal tumor, Squamous cellcarcinoma, Stomach cancer, Synovial sarcoma, T-cell lymphoma),Testicular cancer, Throat cancer, Thymoma/thymic carcinoma, Thyroidcancer, Tongue cancer, Tonsil cancer, Transitional cell cancer,Transitional cell cancer, Transitional cell cancer, Triple-negativebreast cancer, Tubal cancer, Tubular carcinoma, Ureteral cancer,Ureteral cancer, Urethral cancer, Uterine adenocarcinoma, Uterinecancer, Uterine sarcoma, Vaginal cancer, and Vulvar cancer.

49. The use of any one of embodiments 47 and 48, wherein the disease,disorder, or condition is an autoimmunity selected from the groupconsisting of: Autoimmune disseminated encephalomyelitis, Autoimmuneinner ear disease, Batten disease/Neuronal Ceroid Lipofuscinoses,Chronic inflammatory demyelinating polyneuropathy, Encephalitislethargica, Anti-basal ganglia, Guillain-Barré syndrome, Hashimoto'sEncephalopathy, Anti-TPO, Isaac's syndrome/acquired neuromyotonia,Miller Fisher syndrome Morvan's syndrome, Multiple sclerosis, Myastheniagravis, Narcolepsy PANDAS, Rasmussen's encephalitis, Stiff-personsyndrome, Vogt-Koyanagi-Harada syndrome, Addison's disease, Autoimmunehypoparathyroidism, Autoimmune hypophysitis, Autoimmune oophoritis,Autoimmune orchitis, Autoimmune polyglandular syndrome I (APECED),Autoimmune polyglandular syndrome II, Autoimmune polyglandular syndromeIII, Diabetes mellitus, type 1, Graves' disease, Hashimoto's autoimmunethyroiditis, Immunodysregulation, polyendocrinopathy, enteropathy,X-linked, Autoimmune hepatitis type 1, Autoimmune hepatitis type 2,Autoimmune pancreatitis, Coeliac disease, Crohn's disease, Perniciousanemia/atrophic gastritis, Primary biliary cirrhosis, Primary sclerosingcholangitis, Ulcerative colitis, Acquired hemophilia A, Antiphospholipidsyndrome, Autoimmune hemolytic anemia, Autoimmune lymphoproliferativesyndrome, Autoimmune neutropenia, Evans syndrome, Felty's syndrome,Immune thrombocytopenic purpura, Polymyositis/dermatomyositis, Relapsingpolychondritis, Rheumatoid arthritis, Still's disease, Alopecia areata,Bullous pemphigoid, Cicatricial pemphigoid, Dermatitis herpetiformis,Discoid lupus erythematosus, Epidermolysis bullosa acquisita, Linearmorphea, Pemphigus foliaceus, Pemphigus vulgaris, Vitiligo, Behçetdisease, Churg-Strauss syndrome, Cogan's syndrome, CREST syndrome,Anti-fibrillarin, Essential mixed cryoglobulinemia, Mixed connectivetissue disease, POEMS syndrome, Scleroderma, Sjögren's syndrome,Systemic lupus erythematosus, Erythema elevatum diutinum, Kawasakidisease, Microscopic polyangiitis, Polyarteritis nodosa, Rheumaticfever, Takayasu arteritis Temporal arteritis, Wegener's granulomatosis,HLA-B27-associated acute anterior uveitis, Sympathetic ophthalmia, andGoodpasture's disease.

50. A chimeric platelet receptor comprising:

(a) a first region comprising at least a portion of a domain of an ITAMreceptor; and

b) a second region comprising region selected from the group consistingof: (i) a linker or targeting domain encoded by a nucleic acid sequenceselected from the group consisting of SEQ ID NO: 48-51; (ii) at least aportion of a protein selected from the group consisting of: myelinoligodendrocyte glycoprotein (MOG), glutamic acid decarboxylase 2(GAD65), myelin associated glycoprotein (MAG), peripheral myelin protein22 (PMP22), thyroid peroxidase (TPO), voltage-gated potassium channel(VGKC), proteolipid protein (PLP), acetylcholine receptor (AChR),tribbles pseudokinase 2 (TRIB2), N-methyl-D-aspartate (NMDA)-typeglutamate receptor (GluR), glutamate decarboxylase 2 (GAD2), Armadillorepeat containing 9 (ARMC9), Cytochrome P450 Family 21 Subfamily AMember 2 (CYP21 A2), calcium sensing receptor (CASR), nuclearautoantigenic sperm protein (NASP), insulin, thyroid stimulating hormonereceptor (TSHR), thyroperoxidase, asioglycoprotein receptor, CytochromeP450 Family 2 Subfamily D Member 6 (CYP2D6), lactoferrin (LF), tissuetrans-glutaminase (TTG), H/K ATP-ase, Factor XIII (F8),beta2-glycoprotein I (Beta2-GPI), erythrocyte I/I, B2 integrin (ITGB2),granulocyte-colony stimulating factor (G-CSF), glycoprotein (GP)IIb/IIa, collagen II (COLII), fibrinogen (FBG) pia, myeloperoxidase(MPO), cardiac myosin (CYO), proteinase 3 (PRTN3), trichohyalin (TCHH),bullous pemphigoid associated (BP), glycoprotein 1 (GPI), laminin-332(LM332), transglutaminase (TGM), type VII collagen (COLVII), P80 Coilin(COIL), Desmoglein I (DSG1), Desmoglein III (DSG3), SRY-Box 10 (SOX10),small nuclear ribonucleoprotein U1 subunit (70SNRNP70), S-antigen (SAG),and Collagen alpha-3(IV) chain (α3(IV)NC1 collagen); (iii) at least aportion of an antibody selected from the group consisting of; 3F8, 8H9,Abagovomab, Abciximab, Abituzumab, Abrezekimab, Abrilumab, Actoxumab,Adalimumab, Adecatumumab, Atidortoxumab, Aducanumab, Afasevikumab,Afelimomab, Alacizumab pego, Alemtuzumab, Alirocumab, Altumomabpentetate, Amatuximab, Anatumomab mafenatox, Andecaliximab, Anetumabravtansine, Anifrolumab, Anrukinzumab, Apolizumab, Aprutumab ixadotin,Arcitumomab, Ascrinvacumab, Aselizumab, Atezolizumab, Atinumab,Atorolimumab, Avelumab, Azintuxizumab vedotin, Bapineuzumab,Basiliximab, Bavituximab, BCD-100, Bectumomab, Begelomab, Belantamabmafodotin, Belimumab, Bemarituzumab, Benralizumab, Berlimatoxumab,Bermekimab, Bersanlimab, Bertilimumab, Besilesomab, Bevacizumab,Bezlotoxumab, Biciromab, Bimagrumab, Bimekizumab, Birtamimab,Bivatuzumab mertansine, Bleselumab, Blinatumomab, Blontuvetmab,Blosozumab, Bococizumab, Brazikumab, Brentuximab vedotin, Briakinumab,Brodalumab, Brolucizumab, Brontictuzumab, Burosumab, Cabiralizumab,Camidanlumab tesirine, Camrelizumab, Canakinumab, Cantuzumab mertansine,Cantuzumab ravtansine, Caplacizumab, Capromab pendetide, Carlumab,Carotuximab, Catumaxomab, cBR96-doxorubicin immunoconjugate,Cedelizumab, Cemiplimab, Cergutuzumab amunaleukin, Certolizumab pegol,Cetrelimab, Cetuximab, Cibisatamab, Cirmtuzumab, Citatuzumab bogatox,Cixutumumab, Clazakizumab, Clenoliximab, Clivatuzumab tetraxetan,Codrituzumab, Cofetuzumab pelidotin, Coltuximab ravtansine, Conatumumab,Concizumab, Cosfroviximab, Crenezumab, Crizanlizumab, Crotedumab,CR6261, Cusatuzumab, Dacetuzumab, Daclizumab, Dalotuzumab, Dapirolizumabpegol, Daratumumab, Dectrekumab, Demcizumab, Denintuzumab mafodotin,Denosumab, Depatuxizumab mafodotin, Derlotuximab biotin, Detumomab,Dezamizumab, Dinutuximab, Diridavumab, Domagrozumab, Dorlimomab aritox,Dostarlimab, Drozitumab, DS-8201, Duligotuzumab, Dupilumab, Durvalumab,Dusigitumab, Duvortuxizumab, Ecromeximab, Eculizumab, Edobacomab,Edrecolomab, Efalizumab, Efungumab, Eldelumab, Elezanumab, Elgemtumab,Elotuzumab, Elsilimomab, Emactuzumab, Emapalumab, Emibetuzumab,Emicizumab, Enapotamab vedotin, Enavatuzumab, Enfortumab vedotin,Enlimomab pegol, Enoblituzumab, Enokizumab, Enoticumab, Ensituximab,Epitumomab cituxetan, Epratuzumab, Eptinezumab, Erenumab, Erlizumab,Ertumaxomab, Etaracizumab, Etigilimab, Etrolizumab, Evinacumab,Evolocumab, Exbivirumab, Fanolesomab, Faralimomab, Faricimab,Farletuzumab, Fasinumab, FBTA05, Felvizumab, Fezakinumab, Fibatuzumab,Ficlatuzumab, Figitumumab, Firivumab, Flanvotumab, Fletikumab,Flotetuzumab, Fontolizumab, Foralumab, Foravirumab, Fremanezumab,Fresolimumab, Frovocimab, Frunevetmab, Fulranumab, Futuximab,Galcanezumab, Galiximab, Gancotamab, Ganitumab, Gantenerumab,Gatipotuzumab, Gavilimomab, Gedivumab, Gemtuzumab ozogamicin,Gevokizumab, Gilvetmab, Gimsilumab, Girentuximab, Glembatumumab vedotin,Golimumab, Gomiliximab, Gosuranemab, Guselkumab, Ianalumab, Ibalizumab,IBI308, Ibritumomab tiuxetan, Icrucumab, Idarucizumab, Ifabotuzumab,Igovomab, Iladatuzumab vedotin, IMAB362, Imalumab, Imaprelimab,Imciromab, Imgatuzumab, Inclacumab, Indatuximab ravtansine, Indusatumabvedotin, Inebilizumab, Infliximab, Intetumumab, Inolimomab, Inotuzumabozogamicin, Ipilimumab, Iomab-B, Iratumumab, Isatuximab, Iscalimab,Istiratumab, Itolizumab, Ixekizumab, Keliximab, Labetuzumab,Lacnotuzumab, Ladiratuzumab vedotin, Lampalizumab, Lanadelumab,Landogrozumab, Laprituximab emtansine, Larcaviximab, Lebrikizumab,Lemalesomab, Lendalizumab, Lenvervimab, Lenzilumab, Lerdelimumab,Leronlimab, Lesofavumab, Letolizumab, Lexatumumab, Libivirumab,Lifastuzumab vedotin, Ligelizumab, Loncastuximab tesirine, Losatuxizumabvedotin, Lilotomab satetraxetan, Lintuzumab, Lirilumab, Lodelcizumab,Lokivetmab, Lorvotuzumab mertansine, Lucatumumab, Lulizumab pegol,Lumiliximab, Lumretuzumab, Lupartumab amadotin, Lutikizumab,Mapatumumab, Margetuximab, Marstacimab, Maslimomab, Mavrilimumab,Matuzumab, Mepolizumab, Metelimumab, Milatuzumab, Minretumomab,Mirikizumab, Mirvetuximab soravtansine, Mitumomab, Modotuximab,Mogamulizumab, Monalizumab, Morolimumab, Mosunetuzumab, Motavizumab,Moxetumomab pasudotox, Muromonab-CD3, Nacolomab tafenatox, Namilumab,Naptumomab estafenatox, Naratuximab emtansine, Narnatumab, Natalizumab,Navicixizumab, Navivumab, Naxitamab, Nebacumab, Necitumumab,Nemolizumab, NEOD001, Nerelimomab, Nesvacumab, Netakimab, Nimotuzumab,Nirsevimab, Nivolumab, Nofetumomab merpentan, Obiltoxaximab,Obinutuzumab, Ocaratuzumab, Ocrelizumab, Odulimomab, Ofatumumab,Olaratumab, Oleclumab, Olendalizumab, Olokizumab, Omalizumab,Omburtamab, OMS721, Onartuzumab, Ontuxizumab, Onvatilimab, Opicinumab,Oportuzumab monatox, Oregovomab, Orticumab, Otelixizumab, Otilimab,Otlertuzumab, Oxelumab, Ozanezumab, Ozoralizumab, Pagibaximab,Palivizumab, Pamrevlumab, Panitumumab, Pankomab, Panobacumab,Parsatuzumab, Pascolizumab, Pasotuxizumab, Pateclizumab, Patritumab,PDR001, Pembrolizumab, Pemtumomab, Perakizumab, Pertuzumab, Pexelizumab,Pidilizumab, Pinatuzumab vedotin, Pintumomab, Placulumab, Plozalizumab,Pogalizumab, Polatuzumab vedotin, Ponezumab, Porgaviximab, Prasinezumab,Prezalizumab, Priliximab, Pritoxaximab, Pritumumab, PRO 140, Quilizumab,Racotumomab, Radretumab, Rafivirumab, Ralpancizumab, Ramucirumab,Ranevetmab, Ranibizumab, Raxibacumab, Ravagalimab, Ravulizumab,Refanezumab, Regavirumab, Relatlimab, Remtolumab, Reslizumab,Rilotumumab, Rinucumab, Risankizumab, Rituximab, Rivabazumab pegol,Robatumumab, Rmab, Roledumab, Romilkimab, Romosozumab, Rontalizumab,Rosmantuzumab, Rovalpituzumab tesirine, Rovelizumab, Rozanolixizumab,Ruplizumab, SA237, Sacituzumab govitecan, Samalizumab, Samrotamabvedotin, Sarilumab, Satralizumab, Satumomab pendetide, Secukinumab,Selicrelumab, Seribantumab, Setoxaximab, Setrusumab, Sevirumab,Sibrotuzumab, SGN-CD19A, SHP647, Sifalimumab, Siltuximab, Simtuzumab,Siplizumab, Sirtratumab vedotin, Sirukumab, Sofituzumab vedotin,Solanezumab, Solitomab, Sonepcizumab, Sontuzumab, Spartalizumab,Stamulumab, Sulesomab, Suptavumab, Sutimlimab, Suvizumab, Suvratoxumab,Tabalumab, Tacatuzumab tetraxetan, Tadocizumab, Talacotuzumab,Talizumab, Tamtuvetmab, Tanezumab, Taplitumomab paptox, Tarextumab,Tavolimab, Tefibazumab, Telimomab aritox, Telisotuzumab vedotin,Tenatumomab, Teneliximab, Teplizumab, Tepoditamab, Teprotumumab,Tesidolumab, Tetulomab, Tezepelumab, TGN1412, Tibulizumab,Tildrakizumab, Tigatuzumab, Timigutuzumab, Timolumab, Tiragotumab,Tislelizumab, Tisotumab vedotin, TNX-650, Tocilizumab, Tomuzotuximab,Toralizumab, Tosatoxumab, Tositumomab, Tovetumab, Tralokinumab,Trastuzumab, Trastuzumab emtansine, TRBS07, Tregalizumab, Tremelimumab,Trevogrumab, Tucotuzumab celmoleukin, Tuvirumab, Ublituximab,Ulocuplumab, Urelumab, Urtoxazumab, Ustekinumab, Utomilumab,Vadastuximab talirine, Vanalimab, Vandortuzumab vedotin, Vantictumab,Vanucizumab, Vapaliximab, Varisacumab, Varlilumab, Vatelizumab,Vedolizumab, Veltuzumab, Vepalimomab, Vesencumab, Visilizumab,Vobarilizumab, Volociximab, Vonlerolizumab, Vopratelimab, Vorsetuzumabmafodotin, Votumumab, Vunakizumab, Xentuzumab, XMAB-5574, Zalutumumab,Zanolimumab, Zatuximab, Zenocutuzumab, Ziralimumab, Zolbetuximab,Zolimomab aritox; and (iv) a major histocompatibility complex (MHC)class 1 receptor or a major histocompatibility complex (MHC) class 2receptor, wherein the MHC class 1 receptor is bound to a peptide derivedfrom a tumor antigen, a neoantigen, or an autoantigen or the MHC class 2receptor is bound to a peptide derived from a tumor antigen, aneoantigen, or an autoantigen.

51. A therapeutic delivery system comprising:

(a) an engineered platelet presenting the chimeric platelet receptor ofany of embodiments 1-5 or 50, wherein the engineered platelet has beenproduced through genetic modification of a progenitor megakaryocyte tobe non-thrombogenic and non-immunogenic; and

(b) at least one therapeutic agent selected from the group consistingof: a toxin, a protein, a small molecule drug, and a nucleic acidpackaged within a vesicle inside the platelet.

i) wherein the therapeutic agent is the nucleic acid or the protein,loading occurs through expression in a progenitor megakaryocyte, or

ii) wherein the therapeutic agent is loaded by incubation of theengineered platelet with the therapeutic agent.

The invention claimed is:
 1. An engineered megakaryocyte or megakaryocyte progenitor cell line that produces platelets with reduced thrombogenic potential, comprising a disruption of or deletion of at least two genes encoding: (a) a protein involved in recognition of primary stimuli of thrombus formation selected from the group consisting of: GPIb/V/IX, GPVI (GP6), ITGA2B, CLEC2, integrins α_(IIb)β₃, α₂β₁, α₅β₁ and/or α₆β₁; (b) a protein involved in recognition of secondary mediators of thrombus formation selected from the group consisting of Par1, Par4, P2Y12, GPIb/V/IX, the Thromboxane receptor (TBXA2R), P2Y1, P2X1 and/or integrin α_(IIb)β₃; and/or (c) a protein involved in the release of secondary mediators of thrombus formation selected from the group consisting of Cox1, HPS and/or thromboxane-A synthase (TBXAS1).
 2. The engineered megakaryocyte or megakaryocyte progenitor cell line of claim 1 comprising a disruption or deletion of: (a) a gene expressing GPVI or ITGA2B; (b) a gene expressing Par1, Par4 or P2Y12; and (c) a gene expressing Cox1 or HPS.
 3. The engineered megakaryocyte or megakaryocyte progenitor cell line of claim 1 wherein GPVI, ITGA2B, Par1, Par4, P2Y12, Cox1 and HPS genes are disrupted or deleted.
 4. The engineered megakaryocyte or megakaryocyte progenitor cell line of claim 1 that produces platelets that exhibit (a) reduced clotting potential, (b) reduced recruitment by other activated platelets, and/or (c) reduced ability to recruit other platelets.
 5. The engineered megakaryocyte or megakaryocyte progenitor cell line of claim 2 that produces platelets that exhibit (a) reduced clotting potential, (b) reduced recruitment by other activated platelets, and/or (c) reduced ability to recruit other platelets.
 6. The engineered megakaryocyte or megakaryocyte progenitor cell line thereof of claim 1, wherein the disruption of said gene expression comprises contacting the megakaryocyte or megakaryocyte progenitor cell line with a gene-specific RNA interference construct (RNAi), small interfering RNA (siRNA), microRNA (miRNA), or short hairpin RNA (shRNA), thereby disrupting said gene expression.
 7. The engineered megakaryocyte or megakaryocyte progenitor cell line thereof of claim 1 comprising a mutation or deletion of at least a fragment of said gene that disrupts said gene expression.
 8. The megakaryocyte or megakaryocyte progenitor cell line of claim 1 further comprising a mutation or deletion in a gene encoding β2 microglobulin that reduces immunogenicity of platelets generated from the megakaryocyte or megakaryocyte progenitor cell line.
 9. The megakaryocyte or megakaryocyte progenitor cell line of claim 1 further comprising a mutation or deletion in a gene encoding an MHC I protein, wherein the megakaryocyte or megakaryocyte progenitor cell line produces platelets with reduced immunogenicity.
 10. The megakaryocyte or megakaryocyte progenitor cell line of claim 1 further comprising a heterologous nucleic acid encoding a therapeutic agent that is a protein, peptide or RNA.
 11. The megakaryocyte or megakaryocyte progenitor cell line of claim 10 wherein the nucleic acid encoding the therapeutic agent further comprises a platelet-specific promoter.
 12. The megakaryocyte or megakaryocyte progenitor cell line of claim 10 wherein the nucleic acid encoding the therapeutic agent further comprises an α-granule localization signal.
 13. An engineered megakaryocyte or megakaryocyte progenitor cell line of claim 1 further comprising a mutation, deletion and/or insertion of a platelet receptor.
 14. A method of using the megakaryocyte or megakaryocyte progenitor cell line of claim 1 to produce platelets with reduced thrombogenic potential, comprising culturing the megakaryocyte or megakaryocyte progenitor cell line under conditions that generate platelets, and collecting the platelets.
 15. The method of claim 14 further comprising incubating the platelets with one or more diagnostic or therapeutic agents to load the platelets with the diagnostic or therapeutic agents. 